Structured Review

TaKaRa enzymes bamhi
Restriction endonuclease analysis of LaolNPV <t>DNA.</t> Restriction endonuclease profiles following digestion of A) genomic DNAs of the seven isolates of LaolNPV collected in alfalfa crops nearby Montpellier with BglII, EcoRI and PstI enzymes, B) genomic DNAs of Chrysodeixis chalcites NPV (Chch), Helicoverpa armigera SNPV (Hear), Spodoptera littoralis (Spli), Spodopera exigua MNPV (Se) and the virus under study (LaolNPV) with <t>BamHI,</t> BglII, EcoRI and PstI, and C) genomic DNAs of Mamestra brassicae MNPV (Mb), Mamestra configurata NPV-A (MacoA), Mamestra configurata NPV-B (MacoB) and LaolNPV with BglII, EcoRI and PstI.
Enzymes Bamhi, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 26844 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/enzymes bamhi/product/TaKaRa
Average 90 stars, based on 26844 article reviews
Price from $9.99 to $1999.99
enzymes bamhi - by Bioz Stars, 2020-09
90/100 stars

Images

1) Product Images from "Lacanobia oleracea nucleopolyhedrovirus (LaolNPV): A new European species of alphabaculovirus with a narrow host range"

Article Title: Lacanobia oleracea nucleopolyhedrovirus (LaolNPV): A new European species of alphabaculovirus with a narrow host range

Journal: PLoS ONE

doi: 10.1371/journal.pone.0176171

Restriction endonuclease analysis of LaolNPV DNA. Restriction endonuclease profiles following digestion of A) genomic DNAs of the seven isolates of LaolNPV collected in alfalfa crops nearby Montpellier with BglII, EcoRI and PstI enzymes, B) genomic DNAs of Chrysodeixis chalcites NPV (Chch), Helicoverpa armigera SNPV (Hear), Spodoptera littoralis (Spli), Spodopera exigua MNPV (Se) and the virus under study (LaolNPV) with BamHI, BglII, EcoRI and PstI, and C) genomic DNAs of Mamestra brassicae MNPV (Mb), Mamestra configurata NPV-A (MacoA), Mamestra configurata NPV-B (MacoB) and LaolNPV with BglII, EcoRI and PstI.
Figure Legend Snippet: Restriction endonuclease analysis of LaolNPV DNA. Restriction endonuclease profiles following digestion of A) genomic DNAs of the seven isolates of LaolNPV collected in alfalfa crops nearby Montpellier with BglII, EcoRI and PstI enzymes, B) genomic DNAs of Chrysodeixis chalcites NPV (Chch), Helicoverpa armigera SNPV (Hear), Spodoptera littoralis (Spli), Spodopera exigua MNPV (Se) and the virus under study (LaolNPV) with BamHI, BglII, EcoRI and PstI, and C) genomic DNAs of Mamestra brassicae MNPV (Mb), Mamestra configurata NPV-A (MacoA), Mamestra configurata NPV-B (MacoB) and LaolNPV with BglII, EcoRI and PstI.

Techniques Used:

2) Product Images from "Genetic Diversity of Porcine Epidemic Diarrhea Virus With a Naturally Occurring Truncated ORF3 Gene Found in Guangxi, China"

Article Title: Genetic Diversity of Porcine Epidemic Diarrhea Virus With a Naturally Occurring Truncated ORF3 Gene Found in Guangxi, China

Journal: Frontiers in Veterinary Science

doi: 10.3389/fvets.2020.00435

Detection and identification of two distinct bands of the ORF3 gene of PEDV in Guangxi. (A) Amplification of ORF3 gene in PEDV positive samples. M 1 : DL 2000 marker; Lane 1: negative control; Lane 2: predicted product (740 bp); Lane 3: predicted product (740 bp) and large genomic deletion (~358 bp); Lane 4: CV777 strain; Lane 5: Zhejiang-08 strain; Lane 6: AJ1102 strain. (B) Identification of the plasmid, pMD-18T-ORF3, by enzyme digestion using PstI and BamHI. M 2 : DL 5000 marker; Lane 1: The solid arrows indicate the predicted products (740 bp); Lane 2: The dashed arrows indicate products of PEDV variants with a large genomic deletion (~358 bp).
Figure Legend Snippet: Detection and identification of two distinct bands of the ORF3 gene of PEDV in Guangxi. (A) Amplification of ORF3 gene in PEDV positive samples. M 1 : DL 2000 marker; Lane 1: negative control; Lane 2: predicted product (740 bp); Lane 3: predicted product (740 bp) and large genomic deletion (~358 bp); Lane 4: CV777 strain; Lane 5: Zhejiang-08 strain; Lane 6: AJ1102 strain. (B) Identification of the plasmid, pMD-18T-ORF3, by enzyme digestion using PstI and BamHI. M 2 : DL 5000 marker; Lane 1: The solid arrows indicate the predicted products (740 bp); Lane 2: The dashed arrows indicate products of PEDV variants with a large genomic deletion (~358 bp).

Techniques Used: Amplification, Marker, Negative Control, Plasmid Preparation

3) Product Images from "Systematic Targeted Integration to Study Albumin Gene Control Elements"

Article Title: Systematic Targeted Integration to Study Albumin Gene Control Elements

Journal: PLoS ONE

doi: 10.1371/journal.pone.0023234

Characterization of integrated constructs. A: Chromosomal location of the targeting cassette in HuH7-9. Maps show representative integration of the 1.2-kb plasmid P. Plasmids may integrate in two orientations (O1, O2) between RELN and ORC5 , respectively, in the same or opposite direction as transcription of both genes (arrows). PCR using primers R1 and F2 (from LoxP), and F1 and R2 (from flanking chromosomal regions), demonstrated that the integrant ends were intact and determined orientation. B: Representative PCR screening of integrants. O1 amplimersare F1 – R1 (302 bp) and F2 – R2 (599 bp). O2 amplimersare F1 – R2 (347 bp) and F2 – R1 (554 bp). C: Representative maps showing predicted Southern blot bands of integrated constructs P and 1+2+P obtained with KpnI (K) and HpaI (H). The hybridization probe is marked as a bar over the promoter-GFP region, and the promoter and known enhancer are marked in black. D: Southern blot mapping of eight integrated constructs. The blots (left, KpnI ; right, HpaI digests) were probed with a 1.2 kb FseI - SmaI segment of plasmid P. The expected band sizes are listed in Table S3 .
Figure Legend Snippet: Characterization of integrated constructs. A: Chromosomal location of the targeting cassette in HuH7-9. Maps show representative integration of the 1.2-kb plasmid P. Plasmids may integrate in two orientations (O1, O2) between RELN and ORC5 , respectively, in the same or opposite direction as transcription of both genes (arrows). PCR using primers R1 and F2 (from LoxP), and F1 and R2 (from flanking chromosomal regions), demonstrated that the integrant ends were intact and determined orientation. B: Representative PCR screening of integrants. O1 amplimersare F1 – R1 (302 bp) and F2 – R2 (599 bp). O2 amplimersare F1 – R2 (347 bp) and F2 – R1 (554 bp). C: Representative maps showing predicted Southern blot bands of integrated constructs P and 1+2+P obtained with KpnI (K) and HpaI (H). The hybridization probe is marked as a bar over the promoter-GFP region, and the promoter and known enhancer are marked in black. D: Southern blot mapping of eight integrated constructs. The blots (left, KpnI ; right, HpaI digests) were probed with a 1.2 kb FseI - SmaI segment of plasmid P. The expected band sizes are listed in Table S3 .

Techniques Used: Construct, Plasmid Preparation, Polymerase Chain Reaction, Southern Blot, Hybridization

Cloning and recombination strategies. A: Rat Alb and its upstream region. Using restriction sites in the pLL1 linker, GFP was combined with Alb123, Region 1 (a NarI – XhoI segment from −13.7 to −3.9 kb), Region 2 (a segment from −3.9 to −0.2 kb, generated by PCR that added terminal FseI and PstI sites), and Alb123 (−0.2 to the transcription start, with terminal PstI and BglII sites) in a series of reporter plasmids. The 16.5-kb Alb gene was cloned by joining a 4.4 kb proximal segment (transcription start to SmaI , amplified by PCR that added an SgfI site) and a 12.1 kb distal segment ( SmaI – EagI ). To insert the Alb123-GFP reporter as an FseI – SmaI segment, the Alb -containing plasmid was cut with FseI and SgfI , and blunted at the latter site. The resulting reporter plasmids ranged from 4–40 kb ( Table S1 ). B: Schematic map of the 290 bp linker showing the restriction sites for locus assembly. C: Mechanism of RMCE. The targeting cassette encodes an HY-TK fusion protein that makes the cell sensitive to Gancyclovir. Integration of reporter constructs in pLL1 or pLL2 occurs via Cre-mediated recombination, and can take place in two orientations due to the inverted arrangement of the LoxP sites [32] . Loss of the HY-TK gene renders the integrant cells resistant to Gancyclovir.
Figure Legend Snippet: Cloning and recombination strategies. A: Rat Alb and its upstream region. Using restriction sites in the pLL1 linker, GFP was combined with Alb123, Region 1 (a NarI – XhoI segment from −13.7 to −3.9 kb), Region 2 (a segment from −3.9 to −0.2 kb, generated by PCR that added terminal FseI and PstI sites), and Alb123 (−0.2 to the transcription start, with terminal PstI and BglII sites) in a series of reporter plasmids. The 16.5-kb Alb gene was cloned by joining a 4.4 kb proximal segment (transcription start to SmaI , amplified by PCR that added an SgfI site) and a 12.1 kb distal segment ( SmaI – EagI ). To insert the Alb123-GFP reporter as an FseI – SmaI segment, the Alb -containing plasmid was cut with FseI and SgfI , and blunted at the latter site. The resulting reporter plasmids ranged from 4–40 kb ( Table S1 ). B: Schematic map of the 290 bp linker showing the restriction sites for locus assembly. C: Mechanism of RMCE. The targeting cassette encodes an HY-TK fusion protein that makes the cell sensitive to Gancyclovir. Integration of reporter constructs in pLL1 or pLL2 occurs via Cre-mediated recombination, and can take place in two orientations due to the inverted arrangement of the LoxP sites [32] . Loss of the HY-TK gene renders the integrant cells resistant to Gancyclovir.

Techniques Used: Clone Assay, Generated, Polymerase Chain Reaction, Amplification, Plasmid Preparation, Construct

4) Product Images from "Retrieval of entire genes from environmental DNA by inverse PCR with pre-amplification of target genes using primers containing locked nucleic acids"

Article Title: Retrieval of entire genes from environmental DNA by inverse PCR with pre-amplification of target genes using primers containing locked nucleic acids

Journal: Environmental Microbiology

doi: 10.1111/j.1462-2920.2007.01518.x

Capillary electrophoreses of IPCR (odd-numbered lanes) and PAI-PCR (even-numbered lanes) products amplified from DNA extracted from horse vermiform appendixes. The electrophoreses followed by the analysis of the electrophoregrams were automatically performed by using an Agilent 2100 Bioanalyser with a DNA 7500 LabChip® kit (TaKaRa-Bio). The results are shown as a gel-like image. L is the ladder obtained with DNA fragment size markers (bp). The green and pink peaks are the lowest 50 bp and highest 10 380 bp markers respectively. The substrates used for IPCR or PAI-PCR were BamHI-digested DNA (lanes 1 and 2), EcoRI-digested DNA (lanes 3 and 4), HindIII-digested DNA (lanes 5 and 6), PstI-digested DNA (lanes 7 and 8) and XbaI-digested DNA (lanes 9 and 10).
Figure Legend Snippet: Capillary electrophoreses of IPCR (odd-numbered lanes) and PAI-PCR (even-numbered lanes) products amplified from DNA extracted from horse vermiform appendixes. The electrophoreses followed by the analysis of the electrophoregrams were automatically performed by using an Agilent 2100 Bioanalyser with a DNA 7500 LabChip® kit (TaKaRa-Bio). The results are shown as a gel-like image. L is the ladder obtained with DNA fragment size markers (bp). The green and pink peaks are the lowest 50 bp and highest 10 380 bp markers respectively. The substrates used for IPCR or PAI-PCR were BamHI-digested DNA (lanes 1 and 2), EcoRI-digested DNA (lanes 3 and 4), HindIII-digested DNA (lanes 5 and 6), PstI-digested DNA (lanes 7 and 8) and XbaI-digested DNA (lanes 9 and 10).

Techniques Used: Polymerase Chain Reaction, Amplification

5) Product Images from "Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection"

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection

Journal: Journal of Virology

doi: 10.1128/JVI.02205-18

High expression levels of OATP1A2 enhance CaHEV attachment to and infection of LMH cells. (A) Determination of CaHEV propagation in CEH, LMH, LMH GFP , and LMH 1A2GFP cells. The infected cells were collected, and total RNA was extracted to detect the negative-strand (top) and positive-strand (bottom) CaHEV ORF3 genes using TaqMan real-time RT-PCR and negative-strand-specific RT-PCR, respectively. The presence and absence of negative-strand CaHEV RNA are indicated by “+” and “−,” respectively. (B and C) CaHEV attachment (B) and infection (C) assays with LMH, LMH 1A2-GFP , LMH GFP , and LMH 1A2-GFP cells treated with siRNA. CaHEV ORF3 RNA was detected by TaqMan real-time RT-PCR. Error bars indicate the SEM. *, P  
Figure Legend Snippet: High expression levels of OATP1A2 enhance CaHEV attachment to and infection of LMH cells. (A) Determination of CaHEV propagation in CEH, LMH, LMH GFP , and LMH 1A2GFP cells. The infected cells were collected, and total RNA was extracted to detect the negative-strand (top) and positive-strand (bottom) CaHEV ORF3 genes using TaqMan real-time RT-PCR and negative-strand-specific RT-PCR, respectively. The presence and absence of negative-strand CaHEV RNA are indicated by “+” and “−,” respectively. (B and C) CaHEV attachment (B) and infection (C) assays with LMH, LMH 1A2-GFP , LMH GFP , and LMH 1A2-GFP cells treated with siRNA. CaHEV ORF3 RNA was detected by TaqMan real-time RT-PCR. Error bars indicate the SEM. *, P  

Techniques Used: Expressing, Infection, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction

Inhibition of avian HEV attachment and infection of LMH 1A2-GFP and CEH cells. (A) Cytotoxicity analysis of the substrates (or inhibitors) at their maximum working concentrations (50 μM, 50 μM, 10 μM, and 10 μM for CDCA, SC, carvedilol, and imatinib, respectively) for LMH 1A2-GFP cells. (B) Detection of mRNA and protein levels of OATP1A2 expression in LMH 1A2-GFP cells treated with different substrates or an inhibitor. Anti-GFP antibody was used to detect the OATP1A2 fusions for Western blotting. (C) Inhibition of CaHEV attachment to and infection of LMH 1A2-GFP cells by the substrates or inhibitor. After preincubation with CDCA, SC, carvedilol, and imatinib, CaHEV attachment (left) and infection (right) assays were evaluated using LMH 1A2-GFP cells. CaHEV ORF3 RNA was quantified by TaqMan real-time RT-PCR. (D) Blocking CaHEV attachment (left) and infection (right) in LMH 1A2-GFP cells by ap237. (E) Anti-GST-1A2 ecto mouse sera blocked CaHEV attachment to (left) and infection of (right) LMH 1A2-GFP cells. sp239 protein and negative serum were used as controls. CaHEV ORF3 RNA was quantified by TaqMan real-time RT-PCR. Error bars indicate the SEM. (F) Inhibition of CaHEV infection of CEH evaluated using imatinib, ap237, and murine anti-GST-1A2 ecto along with the corresponding dimethyl sulfoxide (DMSO), sp239, and negative mouse sera (**, P  
Figure Legend Snippet: Inhibition of avian HEV attachment and infection of LMH 1A2-GFP and CEH cells. (A) Cytotoxicity analysis of the substrates (or inhibitors) at their maximum working concentrations (50 μM, 50 μM, 10 μM, and 10 μM for CDCA, SC, carvedilol, and imatinib, respectively) for LMH 1A2-GFP cells. (B) Detection of mRNA and protein levels of OATP1A2 expression in LMH 1A2-GFP cells treated with different substrates or an inhibitor. Anti-GFP antibody was used to detect the OATP1A2 fusions for Western blotting. (C) Inhibition of CaHEV attachment to and infection of LMH 1A2-GFP cells by the substrates or inhibitor. After preincubation with CDCA, SC, carvedilol, and imatinib, CaHEV attachment (left) and infection (right) assays were evaluated using LMH 1A2-GFP cells. CaHEV ORF3 RNA was quantified by TaqMan real-time RT-PCR. (D) Blocking CaHEV attachment (left) and infection (right) in LMH 1A2-GFP cells by ap237. (E) Anti-GST-1A2 ecto mouse sera blocked CaHEV attachment to (left) and infection of (right) LMH 1A2-GFP cells. sp239 protein and negative serum were used as controls. CaHEV ORF3 RNA was quantified by TaqMan real-time RT-PCR. Error bars indicate the SEM. (F) Inhibition of CaHEV infection of CEH evaluated using imatinib, ap237, and murine anti-GST-1A2 ecto along with the corresponding dimethyl sulfoxide (DMSO), sp239, and negative mouse sera (**, P  

Techniques Used: Inhibition, Infection, Expressing, Western Blot, Quantitative RT-PCR, Blocking Assay

6) Product Images from "A novel technique for measuring variations in DNA copy-number: competitive genomic polymerase chain reaction"

Article Title: A novel technique for measuring variations in DNA copy-number: competitive genomic polymerase chain reaction

Journal: BMC Genomics

doi: 10.1186/1471-2164-8-206

A, B. Validation of the CGP assay using Fmix vs. Fmix ( A ), or Fmix vs. Mmix ( B ). The horizontal axis displays 132 loci of chromosome X and 60 loci of chromosome 17. The vertical axis represents test/reference log 2 fluorescence ratio of each locus. C, D. Sensitivity of CGP for detecting a change in low-level copy number. CGP was performed to determine the copy number of loci in the chromosome X. Fmix DNA (46, XX) was used as a reference DNA for all reactions. The distribution of test/reference fluorescence ratios for the different test samples of chromosome X DNA ( C ). Vertical axis, the percentage of DNAs; horizontal axis, test/reference fluorescence ratio. The doublet line, bold line, solid line, dotted line and dashed line indicate the Mmix/Fmix, Fmix/Fmix, 47, XXX/Fmix, 48, XXXX/Fmix and 49, XXXXX/Fmix fluorescence ratios, respectively. The mean relative ratios of autosomal DNAs (dotted line) and chromosome X DNA (solid line) from each experiment versus the number of X chromosomes ( D ). Mean (± 1 s.d.) fluorescence ratios of chromosome X DNA were as follows; XY versus XX, 0.69 (0.49–0.89); XX versus XX, 1.00 (0.84–1.15); XXX versus XX, 1.37 (1.10–1.64); XXXX versus XX, 1.62 (1.21–2.02); XXXXX versus XX, 2.08 (1.63–2.52). A dotted line for chromosome X and a solid line for autosomal mean fluorescence ratios were fitted using standard regression analysis.
Figure Legend Snippet: A, B. Validation of the CGP assay using Fmix vs. Fmix ( A ), or Fmix vs. Mmix ( B ). The horizontal axis displays 132 loci of chromosome X and 60 loci of chromosome 17. The vertical axis represents test/reference log 2 fluorescence ratio of each locus. C, D. Sensitivity of CGP for detecting a change in low-level copy number. CGP was performed to determine the copy number of loci in the chromosome X. Fmix DNA (46, XX) was used as a reference DNA for all reactions. The distribution of test/reference fluorescence ratios for the different test samples of chromosome X DNA ( C ). Vertical axis, the percentage of DNAs; horizontal axis, test/reference fluorescence ratio. The doublet line, bold line, solid line, dotted line and dashed line indicate the Mmix/Fmix, Fmix/Fmix, 47, XXX/Fmix, 48, XXXX/Fmix and 49, XXXXX/Fmix fluorescence ratios, respectively. The mean relative ratios of autosomal DNAs (dotted line) and chromosome X DNA (solid line) from each experiment versus the number of X chromosomes ( D ). Mean (± 1 s.d.) fluorescence ratios of chromosome X DNA were as follows; XY versus XX, 0.69 (0.49–0.89); XX versus XX, 1.00 (0.84–1.15); XXX versus XX, 1.37 (1.10–1.64); XXXX versus XX, 1.62 (1.21–2.02); XXXXX versus XX, 2.08 (1.63–2.52). A dotted line for chromosome X and a solid line for autosomal mean fluorescence ratios were fitted using standard regression analysis.

Techniques Used: Fluorescence

Detection of MYCN gene amplification in neuroblastoma cell lines in CGP. A. Confirmation of MYCN amplification in neuroblastoma cell lines by Southern hybridization. B. CGP analysis of neuroblastoma cell lines. DNA copy number profiles for chromosome region 2p25.3-2q14.3 (approximately every 1.3 Mbps) containing the MYCN gene were derived from 96 oligonucleotide primers. All 12 neuroblastoma-derived cell lines were examined, but only the SH-SY5Y, TNB-1, NGP and SK-N-BE lines are displayed. Horizontal axis indicates each locus from 2pter (bp), and vertical axis shows test/reference log 2 fluorescence ratio. The MYCN loci are indicated by black circles. C. CGP high-resolution analysis (approximately every 48 kbps) in neuroblastoma cell lines. DNA copy number profiles for chromosome 2p24.2-2p24.3 containing the MYCN gene were derived from 72 oligonucleotide primers and represented 66 loci. All 12 neuroblastoma-derived cell lines were examined, but only NBL-S, SK-N-DZ, TGW and RTBM1 cell lines are displayed. Horizontal axis indicates each locus from 2pter, and vertical axis shows test/reference log 2 fluorescence ratio. The arrow and black circles denote the MYCN loci. D. Summary of the amplified region surrounding the MYCN gene. The vertical axis indicates the amplified loci between 15.5 Mega (M) bp and 17.5 M bp from 2pter. The black bars denote the DNA amplification sites for each cell line. Regions where amplifications were detected in more than 2 spots of the CGP assay were defined as
Figure Legend Snippet: Detection of MYCN gene amplification in neuroblastoma cell lines in CGP. A. Confirmation of MYCN amplification in neuroblastoma cell lines by Southern hybridization. B. CGP analysis of neuroblastoma cell lines. DNA copy number profiles for chromosome region 2p25.3-2q14.3 (approximately every 1.3 Mbps) containing the MYCN gene were derived from 96 oligonucleotide primers. All 12 neuroblastoma-derived cell lines were examined, but only the SH-SY5Y, TNB-1, NGP and SK-N-BE lines are displayed. Horizontal axis indicates each locus from 2pter (bp), and vertical axis shows test/reference log 2 fluorescence ratio. The MYCN loci are indicated by black circles. C. CGP high-resolution analysis (approximately every 48 kbps) in neuroblastoma cell lines. DNA copy number profiles for chromosome 2p24.2-2p24.3 containing the MYCN gene were derived from 72 oligonucleotide primers and represented 66 loci. All 12 neuroblastoma-derived cell lines were examined, but only NBL-S, SK-N-DZ, TGW and RTBM1 cell lines are displayed. Horizontal axis indicates each locus from 2pter, and vertical axis shows test/reference log 2 fluorescence ratio. The arrow and black circles denote the MYCN loci. D. Summary of the amplified region surrounding the MYCN gene. The vertical axis indicates the amplified loci between 15.5 Mega (M) bp and 17.5 M bp from 2pter. The black bars denote the DNA amplification sites for each cell line. Regions where amplifications were detected in more than 2 spots of the CGP assay were defined as "amplification sites". The dashed line represents the loci of NAG, DDX1, MYCN and FAM49A .

Techniques Used: Amplification, Hybridization, Derivative Assay, Fluorescence

7) Product Images from "Production of dumbbell probe through hairpin cleavage-ligation and increasing RCA sensitivity and specificity by circle to circle amplification"

Article Title: Production of dumbbell probe through hairpin cleavage-ligation and increasing RCA sensitivity and specificity by circle to circle amplification

Journal: Scientific Reports

doi: 10.1038/srep29229

Verification and optimization of hairpin (HP) cleavage-ligation. ( A ) Schematic illustration of dumbbell probe (DP) formation by cleavage-ligation of hairpin. ( B ) DP formation analyzed by PAGE. Only after cleavage, hairpin can be ligated into a band-shifted DP, which can be recleaved into hairpin. ( C ) All of the three kinds of cleaved ends produced by EcoRI, PstI and EcoRV are suitable to form DP efficiently. ( D ) Effects of stem length at the end side of the RE recognition sequence on DP formation. NsiI recognition sequence is underlined. ( E ) Effects of stem length at the loop side of the RE recognition sequence on DP formation. EcoRI recognition sequence is underlined.
Figure Legend Snippet: Verification and optimization of hairpin (HP) cleavage-ligation. ( A ) Schematic illustration of dumbbell probe (DP) formation by cleavage-ligation of hairpin. ( B ) DP formation analyzed by PAGE. Only after cleavage, hairpin can be ligated into a band-shifted DP, which can be recleaved into hairpin. ( C ) All of the three kinds of cleaved ends produced by EcoRI, PstI and EcoRV are suitable to form DP efficiently. ( D ) Effects of stem length at the end side of the RE recognition sequence on DP formation. NsiI recognition sequence is underlined. ( E ) Effects of stem length at the loop side of the RE recognition sequence on DP formation. EcoRI recognition sequence is underlined.

Techniques Used: Ligation, Polyacrylamide Gel Electrophoresis, Produced, Sequencing

8) Product Images from "A novel technique for measuring variations in DNA copy-number: competitive genomic polymerase chain reaction"

Article Title: A novel technique for measuring variations in DNA copy-number: competitive genomic polymerase chain reaction

Journal: BMC Genomics

doi: 10.1186/1471-2164-8-206

A, B. Validation of the CGP assay using Fmix vs. Fmix ( A ), or Fmix vs. Mmix ( B ). The horizontal axis displays 132 loci of chromosome X and 60 loci of chromosome 17. The vertical axis represents test/reference log 2 fluorescence ratio of each locus. C, D. Sensitivity of CGP for detecting a change in low-level copy number. CGP was performed to determine the copy number of loci in the chromosome X. Fmix DNA (46, XX) was used as a reference DNA for all reactions. The distribution of test/reference fluorescence ratios for the different test samples of chromosome X DNA ( C ). Vertical axis, the percentage of DNAs; horizontal axis, test/reference fluorescence ratio. The doublet line, bold line, solid line, dotted line and dashed line indicate the Mmix/Fmix, Fmix/Fmix, 47, XXX/Fmix, 48, XXXX/Fmix and 49, XXXXX/Fmix fluorescence ratios, respectively. The mean relative ratios of autosomal DNAs (dotted line) and chromosome X DNA (solid line) from each experiment versus the number of X chromosomes ( D ). Mean (± 1 s.d.) fluorescence ratios of chromosome X DNA were as follows; XY versus XX, 0.69 (0.49–0.89); XX versus XX, 1.00 (0.84–1.15); XXX versus XX, 1.37 (1.10–1.64); XXXX versus XX, 1.62 (1.21–2.02); XXXXX versus XX, 2.08 (1.63–2.52). A dotted line for chromosome X and a solid line for autosomal mean fluorescence ratios were fitted using standard regression analysis.
Figure Legend Snippet: A, B. Validation of the CGP assay using Fmix vs. Fmix ( A ), or Fmix vs. Mmix ( B ). The horizontal axis displays 132 loci of chromosome X and 60 loci of chromosome 17. The vertical axis represents test/reference log 2 fluorescence ratio of each locus. C, D. Sensitivity of CGP for detecting a change in low-level copy number. CGP was performed to determine the copy number of loci in the chromosome X. Fmix DNA (46, XX) was used as a reference DNA for all reactions. The distribution of test/reference fluorescence ratios for the different test samples of chromosome X DNA ( C ). Vertical axis, the percentage of DNAs; horizontal axis, test/reference fluorescence ratio. The doublet line, bold line, solid line, dotted line and dashed line indicate the Mmix/Fmix, Fmix/Fmix, 47, XXX/Fmix, 48, XXXX/Fmix and 49, XXXXX/Fmix fluorescence ratios, respectively. The mean relative ratios of autosomal DNAs (dotted line) and chromosome X DNA (solid line) from each experiment versus the number of X chromosomes ( D ). Mean (± 1 s.d.) fluorescence ratios of chromosome X DNA were as follows; XY versus XX, 0.69 (0.49–0.89); XX versus XX, 1.00 (0.84–1.15); XXX versus XX, 1.37 (1.10–1.64); XXXX versus XX, 1.62 (1.21–2.02); XXXXX versus XX, 2.08 (1.63–2.52). A dotted line for chromosome X and a solid line for autosomal mean fluorescence ratios were fitted using standard regression analysis.

Techniques Used: Fluorescence

Detection of MYCN gene amplification in neuroblastoma cell lines in CGP. A. Confirmation of MYCN amplification in neuroblastoma cell lines by Southern hybridization. B. CGP analysis of neuroblastoma cell lines. DNA copy number profiles for chromosome region 2p25.3-2q14.3 (approximately every 1.3 Mbps) containing the MYCN gene were derived from 96 oligonucleotide primers. All 12 neuroblastoma-derived cell lines were examined, but only the SH-SY5Y, TNB-1, NGP and SK-N-BE lines are displayed. Horizontal axis indicates each locus from 2pter (bp), and vertical axis shows test/reference log 2 fluorescence ratio. The MYCN loci are indicated by black circles. C. CGP high-resolution analysis (approximately every 48 kbps) in neuroblastoma cell lines. DNA copy number profiles for chromosome 2p24.2-2p24.3 containing the MYCN gene were derived from 72 oligonucleotide primers and represented 66 loci. All 12 neuroblastoma-derived cell lines were examined, but only NBL-S, SK-N-DZ, TGW and RTBM1 cell lines are displayed. Horizontal axis indicates each locus from 2pter, and vertical axis shows test/reference log 2 fluorescence ratio. The arrow and black circles denote the MYCN loci. D. Summary of the amplified region surrounding the MYCN gene. The vertical axis indicates the amplified loci between 15.5 Mega (M) bp and 17.5 M bp from 2pter. The black bars denote the DNA amplification sites for each cell line. Regions where amplifications were detected in more than 2 spots of the CGP assay were defined as
Figure Legend Snippet: Detection of MYCN gene amplification in neuroblastoma cell lines in CGP. A. Confirmation of MYCN amplification in neuroblastoma cell lines by Southern hybridization. B. CGP analysis of neuroblastoma cell lines. DNA copy number profiles for chromosome region 2p25.3-2q14.3 (approximately every 1.3 Mbps) containing the MYCN gene were derived from 96 oligonucleotide primers. All 12 neuroblastoma-derived cell lines were examined, but only the SH-SY5Y, TNB-1, NGP and SK-N-BE lines are displayed. Horizontal axis indicates each locus from 2pter (bp), and vertical axis shows test/reference log 2 fluorescence ratio. The MYCN loci are indicated by black circles. C. CGP high-resolution analysis (approximately every 48 kbps) in neuroblastoma cell lines. DNA copy number profiles for chromosome 2p24.2-2p24.3 containing the MYCN gene were derived from 72 oligonucleotide primers and represented 66 loci. All 12 neuroblastoma-derived cell lines were examined, but only NBL-S, SK-N-DZ, TGW and RTBM1 cell lines are displayed. Horizontal axis indicates each locus from 2pter, and vertical axis shows test/reference log 2 fluorescence ratio. The arrow and black circles denote the MYCN loci. D. Summary of the amplified region surrounding the MYCN gene. The vertical axis indicates the amplified loci between 15.5 Mega (M) bp and 17.5 M bp from 2pter. The black bars denote the DNA amplification sites for each cell line. Regions where amplifications were detected in more than 2 spots of the CGP assay were defined as "amplification sites". The dashed line represents the loci of NAG, DDX1, MYCN and FAM49A .

Techniques Used: Amplification, Hybridization, Derivative Assay, Fluorescence

9) Product Images from "The receptor genes PfBMPR1B and PfBAMBI are involved in regulating shell biomineralization in the pearl oyster Pinctada fucata"

Article Title: The receptor genes PfBMPR1B and PfBAMBI are involved in regulating shell biomineralization in the pearl oyster Pinctada fucata

Journal: Scientific Reports

doi: 10.1038/s41598-017-10011-y

Interactions among Bambi, Bmpr1b and members of the TGFβ/BMP signaling pathway in Pinctada fucata detected by a yeast two-hybrid system. The plasmids are cotransformed into the yeast AH109 and spread on the dropout synthetically defined (SD) medium. The white colonies on SD-Leu-Trp medium (SD-2D) indicate that the plasmids are successfully transformed. The blue colonies on SD-Ade-His-Leu-Trp/X-α-Gal medium (SD-4D- X-α-Gal) indicate the presence of an interaction between two proteins, comparable with that of the positive (AD-T + BD-p53), negative (AD-T + BD-Lam) and empty plasmid (AD + BD) controls. The triangle indicates that the yeast is gradient diluted five times with sterilized water before spreading on the medium.
Figure Legend Snippet: Interactions among Bambi, Bmpr1b and members of the TGFβ/BMP signaling pathway in Pinctada fucata detected by a yeast two-hybrid system. The plasmids are cotransformed into the yeast AH109 and spread on the dropout synthetically defined (SD) medium. The white colonies on SD-Leu-Trp medium (SD-2D) indicate that the plasmids are successfully transformed. The blue colonies on SD-Ade-His-Leu-Trp/X-α-Gal medium (SD-4D- X-α-Gal) indicate the presence of an interaction between two proteins, comparable with that of the positive (AD-T + BD-p53), negative (AD-T + BD-Lam) and empty plasmid (AD + BD) controls. The triangle indicates that the yeast is gradient diluted five times with sterilized water before spreading on the medium.

Techniques Used: Transformation Assay, Laser Capture Microdissection, Plasmid Preparation

10) Product Images from "Retrieval of entire genes from environmental DNA by inverse PCR with pre-amplification of target genes using primers containing locked nucleic acids"

Article Title: Retrieval of entire genes from environmental DNA by inverse PCR with pre-amplification of target genes using primers containing locked nucleic acids

Journal: Environmental Microbiology

doi: 10.1111/j.1462-2920.2007.01518.x

Capillary electrophoreses of IPCR (odd-numbered lanes) and PAI-PCR (even-numbered lanes) products amplified from DNA extracted from horse vermiform appendixes. The electrophoreses followed by the analysis of the electrophoregrams were automatically performed by using an Agilent 2100 Bioanalyser with a DNA 7500 LabChip® kit (TaKaRa-Bio). The results are shown as a gel-like image. L is the ladder obtained with DNA fragment size markers (bp). The green and pink peaks are the lowest 50 bp and highest 10 380 bp markers respectively. The substrates used for IPCR or PAI-PCR were BamHI-digested DNA (lanes 1 and 2), EcoRI-digested DNA (lanes 3 and 4), HindIII-digested DNA (lanes 5 and 6), PstI-digested DNA (lanes 7 and 8) and XbaI-digested DNA (lanes 9 and 10).
Figure Legend Snippet: Capillary electrophoreses of IPCR (odd-numbered lanes) and PAI-PCR (even-numbered lanes) products amplified from DNA extracted from horse vermiform appendixes. The electrophoreses followed by the analysis of the electrophoregrams were automatically performed by using an Agilent 2100 Bioanalyser with a DNA 7500 LabChip® kit (TaKaRa-Bio). The results are shown as a gel-like image. L is the ladder obtained with DNA fragment size markers (bp). The green and pink peaks are the lowest 50 bp and highest 10 380 bp markers respectively. The substrates used for IPCR or PAI-PCR were BamHI-digested DNA (lanes 1 and 2), EcoRI-digested DNA (lanes 3 and 4), HindIII-digested DNA (lanes 5 and 6), PstI-digested DNA (lanes 7 and 8) and XbaI-digested DNA (lanes 9 and 10).

Techniques Used: Polymerase Chain Reaction, Amplification

11) Product Images from "Liposome-based DNA carriers may induce cellular stress response and change gene expression pattern in transfected cells"

Article Title: Liposome-based DNA carriers may induce cellular stress response and change gene expression pattern in transfected cells

Journal: BMC Molecular Biology

doi: 10.1186/1471-2199-12-27

Kinetics of Hspa1b promoter induction after transfection with Lipofectine (CAT assay) . Chloramphenicol acetyltransferase (CAT) activity was observed at different time points after transfection with Lipofectine of p950/CAT6 construct into B16F10 cells. Each experimental point was performed in duplicate. AcCM - acetylchloramphenicol, product of the enzymatic assay. CM - non-acetylated chloramphenicol, substrate of the assay.
Figure Legend Snippet: Kinetics of Hspa1b promoter induction after transfection with Lipofectine (CAT assay) . Chloramphenicol acetyltransferase (CAT) activity was observed at different time points after transfection with Lipofectine of p950/CAT6 construct into B16F10 cells. Each experimental point was performed in duplicate. AcCM - acetylchloramphenicol, product of the enzymatic assay. CM - non-acetylated chloramphenicol, substrate of the assay.

Techniques Used: Transfection, Activity Assay, Construct, Enzymatic Assay

12) Product Images from "Interaction of extracellular S100A4 with RAGE prompts prometastatic activation of A375 melanoma cells"

Article Title: Interaction of extracellular S100A4 with RAGE prompts prometastatic activation of A375 melanoma cells

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.12808

Expression and synthesis of S100A4. ( A ) Relative mRNA expression of S100A4 in A375, A2058 and MEL ‐ JUSO cell lysates was analysed by quantitative real‐time RT ‐ PCR . Displayed are the 2 −ΔCt values, representing the S100A4 gene expression normalized to the β‐actin endogenous reference gene (mean ± S.E.M., n ≥ 6, * P
Figure Legend Snippet: Expression and synthesis of S100A4. ( A ) Relative mRNA expression of S100A4 in A375, A2058 and MEL ‐ JUSO cell lysates was analysed by quantitative real‐time RT ‐ PCR . Displayed are the 2 −ΔCt values, representing the S100A4 gene expression normalized to the β‐actin endogenous reference gene (mean ± S.E.M., n ≥ 6, * P

Techniques Used: Expressing, Quantitative RT-PCR

Synthesis of RAGE and S100A4‐mediated NF ‐κB p65 activation in melanoma cells. ( A ) Representative Western blot shows RAGE protein synthesis in A375, A375‐hS100A4, and A375‐ hRAGE cell lysates. β‐actin was used as loading control. ( B ) NF ‐κB p65 activation in nuclear extracts of wild type A375, wild type A375 cells re‐treated with culture medium of A375‐hS100A4 cells, and A375‐hS100A4 cells (mean ± S.E.M., n = 4). ( C ) Representative Western blot shows S100A4‐mediated up‐regulation of RAGE protein synthesis in wild type A375 cells re‐treated with culture medium of A375‐hS100A4 cells and A375‐hS100A4 cells compared to wild type A375 cells. β‐actin was used as loading control. ( D ) Densitometric analysis of RAGE expression in wild type A375, wild type A375 cells re‐treated with culture medium of A375‐hS100A4 cells, and A375‐ hS 100A4 cells related to β‐actin expression (mean ± S.E.M., n = 3, * P
Figure Legend Snippet: Synthesis of RAGE and S100A4‐mediated NF ‐κB p65 activation in melanoma cells. ( A ) Representative Western blot shows RAGE protein synthesis in A375, A375‐hS100A4, and A375‐ hRAGE cell lysates. β‐actin was used as loading control. ( B ) NF ‐κB p65 activation in nuclear extracts of wild type A375, wild type A375 cells re‐treated with culture medium of A375‐hS100A4 cells, and A375‐hS100A4 cells (mean ± S.E.M., n = 4). ( C ) Representative Western blot shows S100A4‐mediated up‐regulation of RAGE protein synthesis in wild type A375 cells re‐treated with culture medium of A375‐hS100A4 cells and A375‐hS100A4 cells compared to wild type A375 cells. β‐actin was used as loading control. ( D ) Densitometric analysis of RAGE expression in wild type A375, wild type A375 cells re‐treated with culture medium of A375‐hS100A4 cells, and A375‐ hS 100A4 cells related to β‐actin expression (mean ± S.E.M., n = 3, * P

Techniques Used: Activation Assay, Western Blot, Expressing

Influence of S100A4 overexpression on cellular growth in vitro and in vivo . ( A ) Cell growth was evaluated for wild type A375 and A375‐hS100A4 cells (mean ± S.E.M., n ≥ 9, * P
Figure Legend Snippet: Influence of S100A4 overexpression on cellular growth in vitro and in vivo . ( A ) Cell growth was evaluated for wild type A375 and A375‐hS100A4 cells (mean ± S.E.M., n ≥ 9, * P

Techniques Used: Over Expression, In Vitro, In Vivo

Regulation of cell invasion by S100A4‐ RAGE interaction. Relative invasion of wild type A375, A375‐hS100A4, and A375‐ hRAGE cells (untreated, fivefold molar excess (150 ng/ml) of sRAGE , fivefold (150 ng/ml; in A375 cells) or 100‐fold molar excess (3000 ng/ml) of S100A4 (in A375‐ hRAGE cells), S100A4‐si RNA , and blocking Anti‐ RAGE antibody) are shown. Invasion rate of untreated control cells were set as 100% (mean ± S.E.M., n ≥ 3, * P
Figure Legend Snippet: Regulation of cell invasion by S100A4‐ RAGE interaction. Relative invasion of wild type A375, A375‐hS100A4, and A375‐ hRAGE cells (untreated, fivefold molar excess (150 ng/ml) of sRAGE , fivefold (150 ng/ml; in A375 cells) or 100‐fold molar excess (3000 ng/ml) of S100A4 (in A375‐ hRAGE cells), S100A4‐si RNA , and blocking Anti‐ RAGE antibody) are shown. Invasion rate of untreated control cells were set as 100% (mean ± S.E.M., n ≥ 3, * P

Techniques Used: Blocking Assay

Regulation of cell migration by S100A4‐ RAGE interaction. Relative migration of wild type A375, A375‐hS100A4, and A375‐ hRAGE cells (untreated, fivefold molar excess (150 ng/ml) of sRAGE , fivefold (150 ng/ml; in A375 cells) or 100‐fold molar excess (3000 ng/ml) of S100A4 (in A375‐ hRAGE cells), S100A4‐si RNA , and blocking Anti‐ RAGE antibody) are shown. Migration rate of untreated control cells were set as 100% (mean ± S.E.M., n ≥ 3, * P
Figure Legend Snippet: Regulation of cell migration by S100A4‐ RAGE interaction. Relative migration of wild type A375, A375‐hS100A4, and A375‐ hRAGE cells (untreated, fivefold molar excess (150 ng/ml) of sRAGE , fivefold (150 ng/ml; in A375 cells) or 100‐fold molar excess (3000 ng/ml) of S100A4 (in A375‐ hRAGE cells), S100A4‐si RNA , and blocking Anti‐ RAGE antibody) are shown. Migration rate of untreated control cells were set as 100% (mean ± S.E.M., n ≥ 3, * P

Techniques Used: Migration, Blocking Assay

Effect of S100A4 overexpression on cell adhesion, motility, migration, and invasion in A375 cells. Relative adhesion ( A ), relative scratch widths ( B ), relative migration ( C ), or relative invasive capability ( D ) of A375 and A375‐hS100A4 cells was measured after 24 hrs. Adhesion, migration, and invasion rate of untreated control cells were set as 100%. Initial scratch width was set as 100% (mean ± S.E.M., n = 3, * P
Figure Legend Snippet: Effect of S100A4 overexpression on cell adhesion, motility, migration, and invasion in A375 cells. Relative adhesion ( A ), relative scratch widths ( B ), relative migration ( C ), or relative invasive capability ( D ) of A375 and A375‐hS100A4 cells was measured after 24 hrs. Adhesion, migration, and invasion rate of untreated control cells were set as 100%. Initial scratch width was set as 100% (mean ± S.E.M., n = 3, * P

Techniques Used: Over Expression, Migration

Regulation of cell motility by S100A4‐ RAGE interaction. Relative scratch widths of wild type A375, A375‐hS100A4, and A375‐ hRAGE cells either ( A ) untreated, treated with BFA , S100A4‐si RNA , and blocking Anti‐ RAGE antibody, or ( B ) treated with fivefold molar excess (150 ng/ml) of sRAGE , fivefold (150 ng/ml; in A375 cells) or 100‐fold molar excess (3000 ng/ml) of S100A4 (in A375‐ hRAGE cells) are shown. Initial scratch width was set as 100% (mean ± S.E.M., n ≥ 3, * P
Figure Legend Snippet: Regulation of cell motility by S100A4‐ RAGE interaction. Relative scratch widths of wild type A375, A375‐hS100A4, and A375‐ hRAGE cells either ( A ) untreated, treated with BFA , S100A4‐si RNA , and blocking Anti‐ RAGE antibody, or ( B ) treated with fivefold molar excess (150 ng/ml) of sRAGE , fivefold (150 ng/ml; in A375 cells) or 100‐fold molar excess (3000 ng/ml) of S100A4 (in A375‐ hRAGE cells) are shown. Initial scratch width was set as 100% (mean ± S.E.M., n ≥ 3, * P

Techniques Used: Blocking Assay

Active secretion of S100A4. ( A ) Extracellular S100A4 was detected after 4 hrs of incubation in cell culture supernatants of A375, A2058, and MEL ‐ JUSO cells via ELISA (mean ± S.E.M., n = 3, * P
Figure Legend Snippet: Active secretion of S100A4. ( A ) Extracellular S100A4 was detected after 4 hrs of incubation in cell culture supernatants of A375, A2058, and MEL ‐ JUSO cells via ELISA (mean ± S.E.M., n = 3, * P

Techniques Used: Incubation, Cell Culture, Enzyme-linked Immunosorbent Assay

Identification of secretion pathway of S100A4. ( A ) Representative Western blots show detection of extracellular S100A4 in concentrated samples of cell culture supernatants of A375 and A375‐hS100A4 cells analysed untreated or after incubation with bafilomycin A1 (Baf), brefeldin A ( BFA ), cytochalasin B (CytB), or nocodazole (Noc) after 4 hrs of incubation. ( B ) Densitometric analysis of Western blots showing detection of extracellular S100A4 in A375 cells after 4 hrs incubation with different inhibitors related to β‐actin expression (mean ± S.E.M., n = 5, * P
Figure Legend Snippet: Identification of secretion pathway of S100A4. ( A ) Representative Western blots show detection of extracellular S100A4 in concentrated samples of cell culture supernatants of A375 and A375‐hS100A4 cells analysed untreated or after incubation with bafilomycin A1 (Baf), brefeldin A ( BFA ), cytochalasin B (CytB), or nocodazole (Noc) after 4 hrs of incubation. ( B ) Densitometric analysis of Western blots showing detection of extracellular S100A4 in A375 cells after 4 hrs incubation with different inhibitors related to β‐actin expression (mean ± S.E.M., n = 5, * P

Techniques Used: Western Blot, Cell Culture, Incubation, Expressing

13) Product Images from "Retrieval of entire genes from environmental DNA by inverse PCR with pre-amplification of target genes using primers containing locked nucleic acids"

Article Title: Retrieval of entire genes from environmental DNA by inverse PCR with pre-amplification of target genes using primers containing locked nucleic acids

Journal: Environmental Microbiology

doi: 10.1111/j.1462-2920.2007.01518.x

Capillary electrophoreses of IPCR (odd-numbered lanes) and PAI-PCR (even-numbered lanes) products amplified from DNA extracted from horse vermiform appendixes. The electrophoreses followed by the analysis of the electrophoregrams were automatically performed by using an Agilent 2100 Bioanalyser with a DNA 7500 LabChip® kit (TaKaRa-Bio). The results are shown as a gel-like image. L is the ladder obtained with DNA fragment size markers (bp). The green and pink peaks are the lowest 50 bp and highest 10 380 bp markers respectively. The substrates used for IPCR or PAI-PCR were BamHI-digested DNA (lanes 1 and 2), EcoRI-digested DNA (lanes 3 and 4), HindIII-digested DNA (lanes 5 and 6), PstI-digested DNA (lanes 7 and 8) and XbaI-digested DNA (lanes 9 and 10).
Figure Legend Snippet: Capillary electrophoreses of IPCR (odd-numbered lanes) and PAI-PCR (even-numbered lanes) products amplified from DNA extracted from horse vermiform appendixes. The electrophoreses followed by the analysis of the electrophoregrams were automatically performed by using an Agilent 2100 Bioanalyser with a DNA 7500 LabChip® kit (TaKaRa-Bio). The results are shown as a gel-like image. L is the ladder obtained with DNA fragment size markers (bp). The green and pink peaks are the lowest 50 bp and highest 10 380 bp markers respectively. The substrates used for IPCR or PAI-PCR were BamHI-digested DNA (lanes 1 and 2), EcoRI-digested DNA (lanes 3 and 4), HindIII-digested DNA (lanes 5 and 6), PstI-digested DNA (lanes 7 and 8) and XbaI-digested DNA (lanes 9 and 10).

Techniques Used: Polymerase Chain Reaction, Amplification

14) Product Images from "Retrieval of entire genes from environmental DNA by inverse PCR with pre-amplification of target genes using primers containing locked nucleic acids"

Article Title: Retrieval of entire genes from environmental DNA by inverse PCR with pre-amplification of target genes using primers containing locked nucleic acids

Journal: Environmental Microbiology

doi: 10.1111/j.1462-2920.2007.01518.x

Capillary electrophoreses of IPCR (odd-numbered lanes) and PAI-PCR (even-numbered lanes) products amplified from DNA extracted from horse vermiform appendixes. The electrophoreses followed by the analysis of the electrophoregrams were automatically performed by using an Agilent 2100 Bioanalyser with a DNA 7500 LabChip® kit (TaKaRa-Bio). The results are shown as a gel-like image. L is the ladder obtained with DNA fragment size markers (bp). The green and pink peaks are the lowest 50 bp and highest 10 380 bp markers respectively. The substrates used for IPCR or PAI-PCR were BamHI-digested DNA (lanes 1 and 2), EcoRI-digested DNA (lanes 3 and 4), HindIII-digested DNA (lanes 5 and 6), PstI-digested DNA (lanes 7 and 8) and XbaI-digested DNA (lanes 9 and 10).
Figure Legend Snippet: Capillary electrophoreses of IPCR (odd-numbered lanes) and PAI-PCR (even-numbered lanes) products amplified from DNA extracted from horse vermiform appendixes. The electrophoreses followed by the analysis of the electrophoregrams were automatically performed by using an Agilent 2100 Bioanalyser with a DNA 7500 LabChip® kit (TaKaRa-Bio). The results are shown as a gel-like image. L is the ladder obtained with DNA fragment size markers (bp). The green and pink peaks are the lowest 50 bp and highest 10 380 bp markers respectively. The substrates used for IPCR or PAI-PCR were BamHI-digested DNA (lanes 1 and 2), EcoRI-digested DNA (lanes 3 and 4), HindIII-digested DNA (lanes 5 and 6), PstI-digested DNA (lanes 7 and 8) and XbaI-digested DNA (lanes 9 and 10).

Techniques Used: Polymerase Chain Reaction, Amplification

15) Product Images from "Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection"

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection

Journal: Journal of Virology

doi: 10.1128/JVI.02205-18

High expression levels of OATP1A2 enhance CaHEV attachment to and infection of LMH cells. (A) Determination of CaHEV propagation in CEH, LMH, LMH GFP , and LMH 1A2GFP cells. The infected cells were collected, and total RNA was extracted to detect the negative-strand (top) and positive-strand (bottom) CaHEV ORF3 genes using TaqMan real-time RT-PCR and negative-strand-specific RT-PCR, respectively. The presence and absence of negative-strand CaHEV RNA are indicated by “+” and “−,” respectively. (B and C) CaHEV attachment (B) and infection (C) assays with LMH, LMH 1A2-GFP , LMH GFP , and LMH 1A2-GFP cells treated with siRNA. CaHEV ORF3 RNA was detected by TaqMan real-time RT-PCR. Error bars indicate the SEM. *, P
Figure Legend Snippet: High expression levels of OATP1A2 enhance CaHEV attachment to and infection of LMH cells. (A) Determination of CaHEV propagation in CEH, LMH, LMH GFP , and LMH 1A2GFP cells. The infected cells were collected, and total RNA was extracted to detect the negative-strand (top) and positive-strand (bottom) CaHEV ORF3 genes using TaqMan real-time RT-PCR and negative-strand-specific RT-PCR, respectively. The presence and absence of negative-strand CaHEV RNA are indicated by “+” and “−,” respectively. (B and C) CaHEV attachment (B) and infection (C) assays with LMH, LMH 1A2-GFP , LMH GFP , and LMH 1A2-GFP cells treated with siRNA. CaHEV ORF3 RNA was detected by TaqMan real-time RT-PCR. Error bars indicate the SEM. *, P

Techniques Used: Expressing, Infection, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction

Inhibition of avian HEV attachment and infection of LMH 1A2-GFP and CEH cells. (A) Cytotoxicity analysis of the substrates (or inhibitors) at their maximum working concentrations (50 μM, 50 μM, 10 μM, and 10 μM for CDCA, SC, carvedilol, and imatinib, respectively) for LMH 1A2-GFP cells. (B) Detection of mRNA and protein levels of OATP1A2 expression in LMH 1A2-GFP cells treated with different substrates or an inhibitor. Anti-GFP antibody was used to detect the OATP1A2 fusions for Western blotting. (C) Inhibition of CaHEV attachment to and infection of LMH 1A2-GFP cells by the substrates or inhibitor. After preincubation with CDCA, SC, carvedilol, and imatinib, CaHEV attachment (left) and infection (right) assays were evaluated using LMH 1A2-GFP cells. CaHEV ORF3 RNA was quantified by TaqMan real-time RT-PCR. (D) Blocking CaHEV attachment (left) and infection (right) in LMH 1A2-GFP cells by ap237. (E) Anti-GST-1A2 ecto mouse sera blocked CaHEV attachment to (left) and infection of (right) LMH 1A2-GFP cells. sp239 protein and negative serum were used as controls. CaHEV ORF3 RNA was quantified by TaqMan real-time RT-PCR. Error bars indicate the SEM. (F) Inhibition of CaHEV infection of CEH evaluated using imatinib, ap237, and murine anti-GST-1A2 ecto along with the corresponding dimethyl sulfoxide (DMSO), sp239, and negative mouse sera (**, P
Figure Legend Snippet: Inhibition of avian HEV attachment and infection of LMH 1A2-GFP and CEH cells. (A) Cytotoxicity analysis of the substrates (or inhibitors) at their maximum working concentrations (50 μM, 50 μM, 10 μM, and 10 μM for CDCA, SC, carvedilol, and imatinib, respectively) for LMH 1A2-GFP cells. (B) Detection of mRNA and protein levels of OATP1A2 expression in LMH 1A2-GFP cells treated with different substrates or an inhibitor. Anti-GFP antibody was used to detect the OATP1A2 fusions for Western blotting. (C) Inhibition of CaHEV attachment to and infection of LMH 1A2-GFP cells by the substrates or inhibitor. After preincubation with CDCA, SC, carvedilol, and imatinib, CaHEV attachment (left) and infection (right) assays were evaluated using LMH 1A2-GFP cells. CaHEV ORF3 RNA was quantified by TaqMan real-time RT-PCR. (D) Blocking CaHEV attachment (left) and infection (right) in LMH 1A2-GFP cells by ap237. (E) Anti-GST-1A2 ecto mouse sera blocked CaHEV attachment to (left) and infection of (right) LMH 1A2-GFP cells. sp239 protein and negative serum were used as controls. CaHEV ORF3 RNA was quantified by TaqMan real-time RT-PCR. Error bars indicate the SEM. (F) Inhibition of CaHEV infection of CEH evaluated using imatinib, ap237, and murine anti-GST-1A2 ecto along with the corresponding dimethyl sulfoxide (DMSO), sp239, and negative mouse sera (**, P

Techniques Used: Inhibition, Infection, Expressing, Western Blot, Quantitative RT-PCR, Blocking Assay

Related Articles

Transfection:

Article Title: Therapeutic role of human hepatocyte growth factor (HGF) in treating hair loss
Article Snippet: .. Materials E. coli DH5α, E. coli JM109 (Promega, WI, USA), BamHI and SalI (TaKaRa, Japan), pTARGET vector (Promega, Fitchburg, WI, USA), T4 DNA ligase (Promega, Fitchburg, WI, USA), TIAN quick Midi Purification Kit, TIANgel Midi Purification Kit, EndoFree Maxi Plasmid Kit (TIANGEN BIOTECH Co., Ltd.), Polymerase Chain Reaction (PCR) primer (Shanghai Sango Biotech Co., Ltd.), human skin fibroblasts (Academy of Military Medical Sciences), EntransterTM-in vivo Transfection Reagent (Engreen), Lipofectamine™ 2000 (Invitrogen), hHGF enzyme-linked immunosorbent assay (ELISA) Kit (R & D), rabbit anti-HGF monoclonal antibody, rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody, rabbit anti-β-catenin monoclonal antibody, Strept Avidin-biotin complex (SABC) kit and goat anti-rabbit IgG horseradish peroxidase (Wuhan Boster Biological Technology Co., Ltd.), β-catenin, HGF receptor (HGF-R) (Beijing Biosynthesis Biotechnology Co., Ltd.), Diaminobenzidine (DAB) and SP-9001 (ZSGB-BIO), Dulbecco’s modified eagle medium (DMEM) (Gibco), fetal bovine serum (FBS) (Hyclone), streptomycin sulfate and penicillin G sodium (Amresco), Boehringer DIG Luminescent Detection Kit (Boehringer–Mannheim), and Kodak X-Omat AR film (Kodak, Rochester) were commercially available and utilized according to manufacture’s protocol. .. The pEGFP-N1-hHGF was previously constructed in our laboratory ( ; ).

Article Title: Development of TEM-1 β-lactamase based protein translocation assay for identification of Anaplasma phagocytophilum type IV secretion system effector proteins
Article Snippet: .. Briefly, DNA fragment encoding full-length APH0215 was amplified by PCR using a pair of primers (APH0215-GFP-F and APH0215-GFP-R) (Table ) and A. phagocytophilum genomic DNA as template, followed by digestion with XhoI and BamHI, and ligation with eukaryotic expression vector pEGFP-N1 (Clontech), leading to the generation of plasmid pAPH0215-GFP. pAPH0215-GFP and pEGFP-N1 were then transfected into HeLa cells by Polyethylenimine MAX in EZ Trans transfection kit according to manufacturer’s instruction (Shanghai Life iLab Biotech Co, Shanghai, China), respectively. .. For IFA, the transfected cells were harvested at 36 h post-transfection, and subjected to fixation with 2% paraformaldehyde in PBS at RT for 30 min and washing with PBS for three times, followed by permeabilization and blocking with PS solution (PBS containing 0.8% BSA and 0.3% saponin) at RT for 10 min. Immunofluorescence labeling was performed using mouse monoclonal antibody against TGN38 (Clone B-6, Santa Cruz Biotechnology) and Alexa Fluor 555-conjugated goat anti-mouse IgG (Invitrogen).

Amplification:

Article Title: Structural and Functional Characterization of the Actin-1 Gene Promoter From the Antheraea pernyi (Lepidoptera: Saturniidae)
Article Snippet: .. The amplified products were digested with HindIII and BamHI (Takara) and the large DNA restriction fragment purified by agarose gel electrophoresis. .. Plasmid pGFP-N2 (Clontech) was similarly digested and the smaller restriction fragment (GFP gene) was gel purified.

Article Title: Efficient Gene Silencing Mediated by Tobacco Rattle Virus in an Emerging Model Plant Physalis
Article Snippet: .. The amplified products and TRV2 were double digested with Nco I and BamH I (TaKaRa), and were then ligated using T4 DNA ligase (TaKaRa) to generate PfPDS -TRV2 , MPF2 -TRV2 , and MPF3 -TRV2 . ..

Article Title: Development of TEM-1 β-lactamase based protein translocation assay for identification of Anaplasma phagocytophilum type IV secretion system effector proteins
Article Snippet: .. Briefly, DNA fragment encoding full-length APH0215 was amplified by PCR using a pair of primers (APH0215-GFP-F and APH0215-GFP-R) (Table ) and A. phagocytophilum genomic DNA as template, followed by digestion with XhoI and BamHI, and ligation with eukaryotic expression vector pEGFP-N1 (Clontech), leading to the generation of plasmid pAPH0215-GFP. pAPH0215-GFP and pEGFP-N1 were then transfected into HeLa cells by Polyethylenimine MAX in EZ Trans transfection kit according to manufacturer’s instruction (Shanghai Life iLab Biotech Co, Shanghai, China), respectively. .. For IFA, the transfected cells were harvested at 36 h post-transfection, and subjected to fixation with 2% paraformaldehyde in PBS at RT for 30 min and washing with PBS for three times, followed by permeabilization and blocking with PS solution (PBS containing 0.8% BSA and 0.3% saponin) at RT for 10 min. Immunofluorescence labeling was performed using mouse monoclonal antibody against TGN38 (Clone B-6, Santa Cruz Biotechnology) and Alexa Fluor 555-conjugated goat anti-mouse IgG (Invitrogen).

Agarose Gel Electrophoresis:

Article Title: Structural and Functional Characterization of the Actin-1 Gene Promoter From the Antheraea pernyi (Lepidoptera: Saturniidae)
Article Snippet: .. The amplified products were digested with HindIII and BamHI (Takara) and the large DNA restriction fragment purified by agarose gel electrophoresis. .. Plasmid pGFP-N2 (Clontech) was similarly digested and the smaller restriction fragment (GFP gene) was gel purified.

Ligation:

Article Title: Development of TEM-1 β-lactamase based protein translocation assay for identification of Anaplasma phagocytophilum type IV secretion system effector proteins
Article Snippet: .. Briefly, DNA fragment encoding full-length APH0215 was amplified by PCR using a pair of primers (APH0215-GFP-F and APH0215-GFP-R) (Table ) and A. phagocytophilum genomic DNA as template, followed by digestion with XhoI and BamHI, and ligation with eukaryotic expression vector pEGFP-N1 (Clontech), leading to the generation of plasmid pAPH0215-GFP. pAPH0215-GFP and pEGFP-N1 were then transfected into HeLa cells by Polyethylenimine MAX in EZ Trans transfection kit according to manufacturer’s instruction (Shanghai Life iLab Biotech Co, Shanghai, China), respectively. .. For IFA, the transfected cells were harvested at 36 h post-transfection, and subjected to fixation with 2% paraformaldehyde in PBS at RT for 30 min and washing with PBS for three times, followed by permeabilization and blocking with PS solution (PBS containing 0.8% BSA and 0.3% saponin) at RT for 10 min. Immunofluorescence labeling was performed using mouse monoclonal antibody against TGN38 (Clone B-6, Santa Cruz Biotechnology) and Alexa Fluor 555-conjugated goat anti-mouse IgG (Invitrogen).

Avidin-Biotin Assay:

Article Title: Therapeutic role of human hepatocyte growth factor (HGF) in treating hair loss
Article Snippet: .. Materials E. coli DH5α, E. coli JM109 (Promega, WI, USA), BamHI and SalI (TaKaRa, Japan), pTARGET vector (Promega, Fitchburg, WI, USA), T4 DNA ligase (Promega, Fitchburg, WI, USA), TIAN quick Midi Purification Kit, TIANgel Midi Purification Kit, EndoFree Maxi Plasmid Kit (TIANGEN BIOTECH Co., Ltd.), Polymerase Chain Reaction (PCR) primer (Shanghai Sango Biotech Co., Ltd.), human skin fibroblasts (Academy of Military Medical Sciences), EntransterTM-in vivo Transfection Reagent (Engreen), Lipofectamine™ 2000 (Invitrogen), hHGF enzyme-linked immunosorbent assay (ELISA) Kit (R & D), rabbit anti-HGF monoclonal antibody, rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody, rabbit anti-β-catenin monoclonal antibody, Strept Avidin-biotin complex (SABC) kit and goat anti-rabbit IgG horseradish peroxidase (Wuhan Boster Biological Technology Co., Ltd.), β-catenin, HGF receptor (HGF-R) (Beijing Biosynthesis Biotechnology Co., Ltd.), Diaminobenzidine (DAB) and SP-9001 (ZSGB-BIO), Dulbecco’s modified eagle medium (DMEM) (Gibco), fetal bovine serum (FBS) (Hyclone), streptomycin sulfate and penicillin G sodium (Amresco), Boehringer DIG Luminescent Detection Kit (Boehringer–Mannheim), and Kodak X-Omat AR film (Kodak, Rochester) were commercially available and utilized according to manufacture’s protocol. .. The pEGFP-N1-hHGF was previously constructed in our laboratory ( ; ).

Purification:

Article Title: Structural and Functional Characterization of the Actin-1 Gene Promoter From the Antheraea pernyi (Lepidoptera: Saturniidae)
Article Snippet: .. The amplified products were digested with HindIII and BamHI (Takara) and the large DNA restriction fragment purified by agarose gel electrophoresis. .. Plasmid pGFP-N2 (Clontech) was similarly digested and the smaller restriction fragment (GFP gene) was gel purified.

Article Title: Therapeutic role of human hepatocyte growth factor (HGF) in treating hair loss
Article Snippet: .. Materials E. coli DH5α, E. coli JM109 (Promega, WI, USA), BamHI and SalI (TaKaRa, Japan), pTARGET vector (Promega, Fitchburg, WI, USA), T4 DNA ligase (Promega, Fitchburg, WI, USA), TIAN quick Midi Purification Kit, TIANgel Midi Purification Kit, EndoFree Maxi Plasmid Kit (TIANGEN BIOTECH Co., Ltd.), Polymerase Chain Reaction (PCR) primer (Shanghai Sango Biotech Co., Ltd.), human skin fibroblasts (Academy of Military Medical Sciences), EntransterTM-in vivo Transfection Reagent (Engreen), Lipofectamine™ 2000 (Invitrogen), hHGF enzyme-linked immunosorbent assay (ELISA) Kit (R & D), rabbit anti-HGF monoclonal antibody, rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody, rabbit anti-β-catenin monoclonal antibody, Strept Avidin-biotin complex (SABC) kit and goat anti-rabbit IgG horseradish peroxidase (Wuhan Boster Biological Technology Co., Ltd.), β-catenin, HGF receptor (HGF-R) (Beijing Biosynthesis Biotechnology Co., Ltd.), Diaminobenzidine (DAB) and SP-9001 (ZSGB-BIO), Dulbecco’s modified eagle medium (DMEM) (Gibco), fetal bovine serum (FBS) (Hyclone), streptomycin sulfate and penicillin G sodium (Amresco), Boehringer DIG Luminescent Detection Kit (Boehringer–Mannheim), and Kodak X-Omat AR film (Kodak, Rochester) were commercially available and utilized according to manufacture’s protocol. .. The pEGFP-N1-hHGF was previously constructed in our laboratory ( ; ).

Enzyme-linked Immunosorbent Assay:

Article Title: Therapeutic role of human hepatocyte growth factor (HGF) in treating hair loss
Article Snippet: .. Materials E. coli DH5α, E. coli JM109 (Promega, WI, USA), BamHI and SalI (TaKaRa, Japan), pTARGET vector (Promega, Fitchburg, WI, USA), T4 DNA ligase (Promega, Fitchburg, WI, USA), TIAN quick Midi Purification Kit, TIANgel Midi Purification Kit, EndoFree Maxi Plasmid Kit (TIANGEN BIOTECH Co., Ltd.), Polymerase Chain Reaction (PCR) primer (Shanghai Sango Biotech Co., Ltd.), human skin fibroblasts (Academy of Military Medical Sciences), EntransterTM-in vivo Transfection Reagent (Engreen), Lipofectamine™ 2000 (Invitrogen), hHGF enzyme-linked immunosorbent assay (ELISA) Kit (R & D), rabbit anti-HGF monoclonal antibody, rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody, rabbit anti-β-catenin monoclonal antibody, Strept Avidin-biotin complex (SABC) kit and goat anti-rabbit IgG horseradish peroxidase (Wuhan Boster Biological Technology Co., Ltd.), β-catenin, HGF receptor (HGF-R) (Beijing Biosynthesis Biotechnology Co., Ltd.), Diaminobenzidine (DAB) and SP-9001 (ZSGB-BIO), Dulbecco’s modified eagle medium (DMEM) (Gibco), fetal bovine serum (FBS) (Hyclone), streptomycin sulfate and penicillin G sodium (Amresco), Boehringer DIG Luminescent Detection Kit (Boehringer–Mannheim), and Kodak X-Omat AR film (Kodak, Rochester) were commercially available and utilized according to manufacture’s protocol. .. The pEGFP-N1-hHGF was previously constructed in our laboratory ( ; ).

Polymerase Chain Reaction:

Article Title: Therapeutic role of human hepatocyte growth factor (HGF) in treating hair loss
Article Snippet: .. Materials E. coli DH5α, E. coli JM109 (Promega, WI, USA), BamHI and SalI (TaKaRa, Japan), pTARGET vector (Promega, Fitchburg, WI, USA), T4 DNA ligase (Promega, Fitchburg, WI, USA), TIAN quick Midi Purification Kit, TIANgel Midi Purification Kit, EndoFree Maxi Plasmid Kit (TIANGEN BIOTECH Co., Ltd.), Polymerase Chain Reaction (PCR) primer (Shanghai Sango Biotech Co., Ltd.), human skin fibroblasts (Academy of Military Medical Sciences), EntransterTM-in vivo Transfection Reagent (Engreen), Lipofectamine™ 2000 (Invitrogen), hHGF enzyme-linked immunosorbent assay (ELISA) Kit (R & D), rabbit anti-HGF monoclonal antibody, rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody, rabbit anti-β-catenin monoclonal antibody, Strept Avidin-biotin complex (SABC) kit and goat anti-rabbit IgG horseradish peroxidase (Wuhan Boster Biological Technology Co., Ltd.), β-catenin, HGF receptor (HGF-R) (Beijing Biosynthesis Biotechnology Co., Ltd.), Diaminobenzidine (DAB) and SP-9001 (ZSGB-BIO), Dulbecco’s modified eagle medium (DMEM) (Gibco), fetal bovine serum (FBS) (Hyclone), streptomycin sulfate and penicillin G sodium (Amresco), Boehringer DIG Luminescent Detection Kit (Boehringer–Mannheim), and Kodak X-Omat AR film (Kodak, Rochester) were commercially available and utilized according to manufacture’s protocol. .. The pEGFP-N1-hHGF was previously constructed in our laboratory ( ; ).

Article Title: Development of TEM-1 β-lactamase based protein translocation assay for identification of Anaplasma phagocytophilum type IV secretion system effector proteins
Article Snippet: .. Briefly, DNA fragment encoding full-length APH0215 was amplified by PCR using a pair of primers (APH0215-GFP-F and APH0215-GFP-R) (Table ) and A. phagocytophilum genomic DNA as template, followed by digestion with XhoI and BamHI, and ligation with eukaryotic expression vector pEGFP-N1 (Clontech), leading to the generation of plasmid pAPH0215-GFP. pAPH0215-GFP and pEGFP-N1 were then transfected into HeLa cells by Polyethylenimine MAX in EZ Trans transfection kit according to manufacturer’s instruction (Shanghai Life iLab Biotech Co, Shanghai, China), respectively. .. For IFA, the transfected cells were harvested at 36 h post-transfection, and subjected to fixation with 2% paraformaldehyde in PBS at RT for 30 min and washing with PBS for three times, followed by permeabilization and blocking with PS solution (PBS containing 0.8% BSA and 0.3% saponin) at RT for 10 min. Immunofluorescence labeling was performed using mouse monoclonal antibody against TGN38 (Clone B-6, Santa Cruz Biotechnology) and Alexa Fluor 555-conjugated goat anti-mouse IgG (Invitrogen).

Expressing:

Article Title: Development of TEM-1 β-lactamase based protein translocation assay for identification of Anaplasma phagocytophilum type IV secretion system effector proteins
Article Snippet: .. Briefly, DNA fragment encoding full-length APH0215 was amplified by PCR using a pair of primers (APH0215-GFP-F and APH0215-GFP-R) (Table ) and A. phagocytophilum genomic DNA as template, followed by digestion with XhoI and BamHI, and ligation with eukaryotic expression vector pEGFP-N1 (Clontech), leading to the generation of plasmid pAPH0215-GFP. pAPH0215-GFP and pEGFP-N1 were then transfected into HeLa cells by Polyethylenimine MAX in EZ Trans transfection kit according to manufacturer’s instruction (Shanghai Life iLab Biotech Co, Shanghai, China), respectively. .. For IFA, the transfected cells were harvested at 36 h post-transfection, and subjected to fixation with 2% paraformaldehyde in PBS at RT for 30 min and washing with PBS for three times, followed by permeabilization and blocking with PS solution (PBS containing 0.8% BSA and 0.3% saponin) at RT for 10 min. Immunofluorescence labeling was performed using mouse monoclonal antibody against TGN38 (Clone B-6, Santa Cruz Biotechnology) and Alexa Fluor 555-conjugated goat anti-mouse IgG (Invitrogen).

Modification:

Article Title: Therapeutic role of human hepatocyte growth factor (HGF) in treating hair loss
Article Snippet: .. Materials E. coli DH5α, E. coli JM109 (Promega, WI, USA), BamHI and SalI (TaKaRa, Japan), pTARGET vector (Promega, Fitchburg, WI, USA), T4 DNA ligase (Promega, Fitchburg, WI, USA), TIAN quick Midi Purification Kit, TIANgel Midi Purification Kit, EndoFree Maxi Plasmid Kit (TIANGEN BIOTECH Co., Ltd.), Polymerase Chain Reaction (PCR) primer (Shanghai Sango Biotech Co., Ltd.), human skin fibroblasts (Academy of Military Medical Sciences), EntransterTM-in vivo Transfection Reagent (Engreen), Lipofectamine™ 2000 (Invitrogen), hHGF enzyme-linked immunosorbent assay (ELISA) Kit (R & D), rabbit anti-HGF monoclonal antibody, rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody, rabbit anti-β-catenin monoclonal antibody, Strept Avidin-biotin complex (SABC) kit and goat anti-rabbit IgG horseradish peroxidase (Wuhan Boster Biological Technology Co., Ltd.), β-catenin, HGF receptor (HGF-R) (Beijing Biosynthesis Biotechnology Co., Ltd.), Diaminobenzidine (DAB) and SP-9001 (ZSGB-BIO), Dulbecco’s modified eagle medium (DMEM) (Gibco), fetal bovine serum (FBS) (Hyclone), streptomycin sulfate and penicillin G sodium (Amresco), Boehringer DIG Luminescent Detection Kit (Boehringer–Mannheim), and Kodak X-Omat AR film (Kodak, Rochester) were commercially available and utilized according to manufacture’s protocol. .. The pEGFP-N1-hHGF was previously constructed in our laboratory ( ; ).

Recombinant:

Article Title: Characterization of the novel In1059 harbouring VIM gene cassette
Article Snippet: .. The plasmids were digested with BamHI (TaKaRa, Dalian, China), and the six integron-harbouring recombinant plasmids were electrophoretically resolved to generate genetic maps. .. To characterize the recombinant plasmid-encoding integrons from the above six isolates, we digested the plasmids with BamHI and ligated the relevant fragments to the pMD19-T cloning vector (TaKaRa, Dalian, China), and then transformed the ligation mixture into E. coli TOP10 host bacteria.

Plasmid Preparation:

Article Title: Dissemination of blaOXA-23 in Acinetobacter spp. in China: Main Roles of Conjugative Plasmid pAZJ221 and Transposon Tn2009
Article Snippet: .. In order to identify plasmid patterns, genomic DNA of isolates with the plasmid-borne bla OXA-23 gene, digested with BamHI or EcoRI (TaKaRa, Japan), was separated and hybridized, as described above. .. Furthermore, genomic DNA digested by ApaI (TaKaRa, Japan) was electrophoresed and then hybridized, as described above, to reveal the chromosomal locations of the bla OXA-23 gene through comparison of hybridization signals with S1-PFGE.

Article Title: Therapeutic role of human hepatocyte growth factor (HGF) in treating hair loss
Article Snippet: .. Materials E. coli DH5α, E. coli JM109 (Promega, WI, USA), BamHI and SalI (TaKaRa, Japan), pTARGET vector (Promega, Fitchburg, WI, USA), T4 DNA ligase (Promega, Fitchburg, WI, USA), TIAN quick Midi Purification Kit, TIANgel Midi Purification Kit, EndoFree Maxi Plasmid Kit (TIANGEN BIOTECH Co., Ltd.), Polymerase Chain Reaction (PCR) primer (Shanghai Sango Biotech Co., Ltd.), human skin fibroblasts (Academy of Military Medical Sciences), EntransterTM-in vivo Transfection Reagent (Engreen), Lipofectamine™ 2000 (Invitrogen), hHGF enzyme-linked immunosorbent assay (ELISA) Kit (R & D), rabbit anti-HGF monoclonal antibody, rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody, rabbit anti-β-catenin monoclonal antibody, Strept Avidin-biotin complex (SABC) kit and goat anti-rabbit IgG horseradish peroxidase (Wuhan Boster Biological Technology Co., Ltd.), β-catenin, HGF receptor (HGF-R) (Beijing Biosynthesis Biotechnology Co., Ltd.), Diaminobenzidine (DAB) and SP-9001 (ZSGB-BIO), Dulbecco’s modified eagle medium (DMEM) (Gibco), fetal bovine serum (FBS) (Hyclone), streptomycin sulfate and penicillin G sodium (Amresco), Boehringer DIG Luminescent Detection Kit (Boehringer–Mannheim), and Kodak X-Omat AR film (Kodak, Rochester) were commercially available and utilized according to manufacture’s protocol. .. The pEGFP-N1-hHGF was previously constructed in our laboratory ( ; ).

Article Title: Development of TEM-1 β-lactamase based protein translocation assay for identification of Anaplasma phagocytophilum type IV secretion system effector proteins
Article Snippet: .. Briefly, DNA fragment encoding full-length APH0215 was amplified by PCR using a pair of primers (APH0215-GFP-F and APH0215-GFP-R) (Table ) and A. phagocytophilum genomic DNA as template, followed by digestion with XhoI and BamHI, and ligation with eukaryotic expression vector pEGFP-N1 (Clontech), leading to the generation of plasmid pAPH0215-GFP. pAPH0215-GFP and pEGFP-N1 were then transfected into HeLa cells by Polyethylenimine MAX in EZ Trans transfection kit according to manufacturer’s instruction (Shanghai Life iLab Biotech Co, Shanghai, China), respectively. .. For IFA, the transfected cells were harvested at 36 h post-transfection, and subjected to fixation with 2% paraformaldehyde in PBS at RT for 30 min and washing with PBS for three times, followed by permeabilization and blocking with PS solution (PBS containing 0.8% BSA and 0.3% saponin) at RT for 10 min. Immunofluorescence labeling was performed using mouse monoclonal antibody against TGN38 (Clone B-6, Santa Cruz Biotechnology) and Alexa Fluor 555-conjugated goat anti-mouse IgG (Invitrogen).

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 89
    TaKaRa restriction enzyme psti
    Restriction Enzyme Psti, supplied by TaKaRa, used in various techniques. Bioz Stars score: 89/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/restriction enzyme psti/product/TaKaRa
    Average 89 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    restriction enzyme psti - by Bioz Stars, 2020-09
    89/100 stars
      Buy from Supplier

    85
    TaKaRa psti enzymes
    Psti Enzymes, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/psti enzymes/product/TaKaRa
    Average 85 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    psti enzymes - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    Image Search Results