Structured Review

Thermo Fisher pcr
Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 75820 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pcr/product/Thermo Fisher
Average 99 stars, based on 75820 article reviews
Price from $9.99 to $1999.99
pcr - by Bioz Stars, 2020-04
99/100 stars

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Related Articles

DNA Extraction:

Article Title: Clinical and Genetic Features of Familial Cold Urticaria: A Report of Three Families
Article Snippet: Genomic DNA was extracted using a blood DNA extraction kit (Tiangen, Biotech Co., Beijing, China). .. The PCR products were sequenced using Big Dye Terminator (Applied Biosystems, Torrance, CA, USA) and ABI PRISM 3730 Xl Genetic Analyzer (Applied Biosystems, Foster City, CA, USA). cDNA of phospholipase C gamma 2 (PLCG2 ) were screened by means of PCR amplification of overlapping segments and then Sanger sequencing.

SYBR Green Assay:

Article Title: Impact of storage conditions on the quality of nucleic acids in paraffin embedded tissues
Article Snippet: .. Four microliters of 1:32 cDNA dilutions served as template for PCR using Power SYBR Green PCR Master Mix (Thermo Fisher Scientific, Wilmington, DE). .. Fragments of human GAPDH with amplicon lengths of 71, 153, 200, 277 and 323 base pair (bp) were amplified as published previously [ ].

Amplification:

Article Title: Impact of storage conditions on the quality of nucleic acids in paraffin embedded tissues
Article Snippet: Four microliters of 1:32 cDNA dilutions served as template for PCR using Power SYBR Green PCR Master Mix (Thermo Fisher Scientific, Wilmington, DE). .. Fragments of human GAPDH with amplicon lengths of 71, 153, 200, 277 and 323 base pair (bp) were amplified as published previously [ ].

Article Title: Clinical and Genetic Features of Familial Cold Urticaria: A Report of Three Families
Article Snippet: .. The PCR products were sequenced using Big Dye Terminator (Applied Biosystems, Torrance, CA, USA) and ABI PRISM 3730 Xl Genetic Analyzer (Applied Biosystems, Foster City, CA, USA). cDNA of phospholipase C gamma 2 (PLCG2 ) were screened by means of PCR amplification of overlapping segments and then Sanger sequencing. .. Human leukocyte antigen (HLA )-A , B , and DRB1 locus-specific amplification was performed using a PCR kit (Qiagen, Hilden, Germany).

Quantitative RT-PCR:

Article Title: Impact of storage conditions on the quality of nucleic acids in paraffin embedded tissues
Article Snippet: Paragraph title: Quantitative RT-PCR (RT-qPCR) ... Four microliters of 1:32 cDNA dilutions served as template for PCR using Power SYBR Green PCR Master Mix (Thermo Fisher Scientific, Wilmington, DE).

Purification:

Article Title: Transmission of Foot-and-Mouth Disease from Persistently Infected Carrier Cattle to Naive Cattle via Transfer of Oropharyngeal Fluid
Article Snippet: 810 bp) were purified using a QIAquick gel extraction and purification kit (Qiagen, Hilden, Germany) and quantified using a Nanodrop 1000 spectrophotometer (Fisher Scientific, Waltham, MA, USA). .. Sequencing of the PCR products was performed using a BigDye Terminator v 3.1 cycle sequencing kit (Applied Biosystems, Foster City, CA, USA), using the same primers as used in the PCRs, according to the manufacturer’s instructions, and ran on an automated DNA sequencer (ABI PRISM 3730 DNA analyzer; Applied Biosystems, Foster City, CA, USA).

Real-time Polymerase Chain Reaction:

Article Title: Impact of storage conditions on the quality of nucleic acids in paraffin embedded tissues
Article Snippet: Four microliters of 1:32 cDNA dilutions served as template for PCR using Power SYBR Green PCR Master Mix (Thermo Fisher Scientific, Wilmington, DE). .. All samples and controls were analyzed in triplicates for 45 cycles of PCR using a MicroAmp fast 96-well reaction plate format on an ABI 7900 Real-Time PCR System (Thermo Fisher Scientific, Wilmington, DE).

Polymerase Chain Reaction:

Article Title: Impact of storage conditions on the quality of nucleic acids in paraffin embedded tissues
Article Snippet: .. Four microliters of 1:32 cDNA dilutions served as template for PCR using Power SYBR Green PCR Master Mix (Thermo Fisher Scientific, Wilmington, DE). .. Fragments of human GAPDH with amplicon lengths of 71, 153, 200, 277 and 323 base pair (bp) were amplified as published previously [ ].

Article Title: Transmission of Foot-and-Mouth Disease from Persistently Infected Carrier Cattle to Naive Cattle via Transfer of Oropharyngeal Fluid
Article Snippet: .. Sequencing of the PCR products was performed using a BigDye Terminator v 3.1 cycle sequencing kit (Applied Biosystems, Foster City, CA, USA), using the same primers as used in the PCRs, according to the manufacturer’s instructions, and ran on an automated DNA sequencer (ABI PRISM 3730 DNA analyzer; Applied Biosystems, Foster City, CA, USA). ..

Article Title: Clinical and Genetic Features of Familial Cold Urticaria: A Report of Three Families
Article Snippet: .. The PCR products were sequenced using Big Dye Terminator (Applied Biosystems, Torrance, CA, USA) and ABI PRISM 3730 Xl Genetic Analyzer (Applied Biosystems, Foster City, CA, USA). cDNA of phospholipase C gamma 2 (PLCG2 ) were screened by means of PCR amplification of overlapping segments and then Sanger sequencing. .. Human leukocyte antigen (HLA )-A , B , and DRB1 locus-specific amplification was performed using a PCR kit (Qiagen, Hilden, Germany).

Random Hexamer Labeling:

Article Title: Transmission of Foot-and-Mouth Disease from Persistently Infected Carrier Cattle to Naive Cattle via Transfer of Oropharyngeal Fluid
Article Snippet: The cDNA synthesis was carried out using Ready-To-Go You-Prime First-Strand beads (GE Healthcare Life Sciences, Uppsala, Sweden) with a mixture of random hexamer primers [pd(N)6] and oligo(dT). .. Sequencing of the PCR products was performed using a BigDye Terminator v 3.1 cycle sequencing kit (Applied Biosystems, Foster City, CA, USA), using the same primers as used in the PCRs, according to the manufacturer’s instructions, and ran on an automated DNA sequencer (ABI PRISM 3730 DNA analyzer; Applied Biosystems, Foster City, CA, USA).

Spectrophotometry:

Article Title: Transmission of Foot-and-Mouth Disease from Persistently Infected Carrier Cattle to Naive Cattle via Transfer of Oropharyngeal Fluid
Article Snippet: 810 bp) were purified using a QIAquick gel extraction and purification kit (Qiagen, Hilden, Germany) and quantified using a Nanodrop 1000 spectrophotometer (Fisher Scientific, Waltham, MA, USA). .. Sequencing of the PCR products was performed using a BigDye Terminator v 3.1 cycle sequencing kit (Applied Biosystems, Foster City, CA, USA), using the same primers as used in the PCRs, according to the manufacturer’s instructions, and ran on an automated DNA sequencer (ABI PRISM 3730 DNA analyzer; Applied Biosystems, Foster City, CA, USA).

Sequencing:

Article Title: Transmission of Foot-and-Mouth Disease from Persistently Infected Carrier Cattle to Naive Cattle via Transfer of Oropharyngeal Fluid
Article Snippet: .. Sequencing of the PCR products was performed using a BigDye Terminator v 3.1 cycle sequencing kit (Applied Biosystems, Foster City, CA, USA), using the same primers as used in the PCRs, according to the manufacturer’s instructions, and ran on an automated DNA sequencer (ABI PRISM 3730 DNA analyzer; Applied Biosystems, Foster City, CA, USA). ..

Article Title: Clinical and Genetic Features of Familial Cold Urticaria: A Report of Three Families
Article Snippet: .. The PCR products were sequenced using Big Dye Terminator (Applied Biosystems, Torrance, CA, USA) and ABI PRISM 3730 Xl Genetic Analyzer (Applied Biosystems, Foster City, CA, USA). cDNA of phospholipase C gamma 2 (PLCG2 ) were screened by means of PCR amplification of overlapping segments and then Sanger sequencing. .. Human leukocyte antigen (HLA )-A , B , and DRB1 locus-specific amplification was performed using a PCR kit (Qiagen, Hilden, Germany).

Gel Extraction:

Article Title: Transmission of Foot-and-Mouth Disease from Persistently Infected Carrier Cattle to Naive Cattle via Transfer of Oropharyngeal Fluid
Article Snippet: 810 bp) were purified using a QIAquick gel extraction and purification kit (Qiagen, Hilden, Germany) and quantified using a Nanodrop 1000 spectrophotometer (Fisher Scientific, Waltham, MA, USA). .. Sequencing of the PCR products was performed using a BigDye Terminator v 3.1 cycle sequencing kit (Applied Biosystems, Foster City, CA, USA), using the same primers as used in the PCRs, according to the manufacturer’s instructions, and ran on an automated DNA sequencer (ABI PRISM 3730 DNA analyzer; Applied Biosystems, Foster City, CA, USA).

Software:

Article Title: Impact of storage conditions on the quality of nucleic acids in paraffin embedded tissues
Article Snippet: Four microliters of 1:32 cDNA dilutions served as template for PCR using Power SYBR Green PCR Master Mix (Thermo Fisher Scientific, Wilmington, DE). .. Results were analyzed using ABI SDS software version 2.3.

Article Title: Clinical and Genetic Features of Familial Cold Urticaria: A Report of Three Families
Article Snippet: The PCR products were sequenced using Big Dye Terminator (Applied Biosystems, Torrance, CA, USA) and ABI PRISM 3730 Xl Genetic Analyzer (Applied Biosystems, Foster City, CA, USA). cDNA of phospholipase C gamma 2 (PLCG2 ) were screened by means of PCR amplification of overlapping segments and then Sanger sequencing. .. The sequencing data were analyzed using SBT engine software (SBT excellerator, Genome Diagnostics B.V., Arnhem, the Netherlands).

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  • 95
    Thermo Fisher quan titative reverse transcriptase polymerase chain reaction qrt pcr
    miR-138 up regulation leads to hTERT suppression in NB4 cells. A: total RNAs from NB4 cells that were transduced with GFP hsa-miR-138-expressing lentiviruses were extracted at 96 h after removal of the lentivirus-containing medium. hTERT mRNA expression was measured by <t>qRT-PCR.</t> Values were normalized to GAPDH . (n=3; *P
    Quan Titative Reverse Transcriptase Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quan titative reverse transcriptase polymerase chain reaction qrt pcr/product/Thermo Fisher
    Average 95 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    quan titative reverse transcriptase polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    95/100 stars
      Buy from Supplier

    99
    Thermo Fisher qrt pcr rna
    MARF1 binds the CCR4-NOT deadenylase complex and cyclin A mRNA. a Co-immunoprecipitation using anti-HA antibody-conjugated beads followed by Western blotting. Ovary lysates expressing 3xHA-MARF1 (MAT67Tub-Gal4 → UASP-3xHA-MARF1) and those from w 1118 negative control were tested. b Co-immunoprecipitation using anti-HA antibody-conjugated beads followed by Western blotting. Ovary lysates expressing 3xHA-tagged MARF1 (MAT67Tub-Gal4 → UASP-3xHA-MARF1 full-length or fragments) and those from w 1118 negative control were tested. Black triangles indicate the detected 3xHA-tagged MARF1 proteins. c Co-immunoprecipitation using ovary lysates from w 1118 and anti-Not1 antibody or negative control IgG-bound protein G beads followed by Western blotting. d Fold enrichment of mRNAs relative to a control gapdh mRNA that were co-immunoprecipitated with MARF1 by anti-HA antibody-conjugated beads and were eluted from the beads normalized by w 1118 negative control, determined by <t>RNA-immunoprecipitation</t> followed by <t>qRT-PCR.</t> Mean ± SD ( n = 3). P -value
    Qrt Pcr Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 285 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/qrt pcr rna/product/Thermo Fisher
    Average 99 stars, based on 285 article reviews
    Price from $9.99 to $1999.99
    qrt pcr rna - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    97
    Thermo Fisher quantitative real time reverse transcription polymerase chain reaction qrt pcr
    Knockdown or suppression of p65 reduces GLS1 expression in HCC cells. ( A ) Western blot shows the knockdown of c-Myc without interfering with GLS1 expression in HepG2 and Hep3B cells. ( B ) Western blot shows that p65 knockdown reduces GLS1 protein levels in both HepG2 and Hep3B cells. ( C ) <t>qRT-PCR</t> indicates that p65 knockdown reduces GLS1 mRNA levels in both HepG2 and Hep3B cells. ** p
    Quantitative Real Time Reverse Transcription Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time reverse transcription polymerase chain reaction qrt pcr/product/Thermo Fisher
    Average 97 stars, based on 44 article reviews
    Price from $9.99 to $1999.99
    quantitative real time reverse transcription polymerase chain reaction qrt pcr - by Bioz Stars, 2020-04
    97/100 stars
      Buy from Supplier

    99
    Thermo Fisher pcr analysis
    Production, purification, and characterization of the INS19 fragment expressed by cgd6_5510 . (A) Electrophoresis analysis of a <t>PCR</t> product from the cgd6_5510 region. Lane M: <t>DNA</t> marker; Lane 1: cgd6_5510 PCR product. (B) The Western blot analysis of the expression of recombinant INS19 in Escherichia coli Rosetta (DE3). Lane M: protein marker; Lane 1: lysate of un-induced bacterial cells transformed with pET28s- cgd6_5510 ; Lane 2: supernatant from lysate of bacterial cells induced by IPTG; Lane 3: inclusion bodies from lysate of bacterial cells induced by IPTG. (C) Purification of recombinant INS19 in inclusion bodies as indicated by results of SDS-PAGE analysis. Lane M: protein marker; Lane 1: proteins from inclusion bodies dissolved in wash buffer containing 1 M urea; Lane 2: proteins from inclusion bodies dissolved in wash buffer containing 2 M urea; Lane 3: proteins from inclusion bodies dissolved in buffer containing 8 M urea; Lane 4: undissolved proteins from inclusion bodies after washing; Lane 5: purified recombinant INS19 after dialysis and ultrafiltration. (D) The Western blot analysis of purified INS19 (Lane 1), C. parvum sporozoite lysate (Lane 2), and C. parvum oocyst lysate (Lane 3) probed with the pre-immune serum (left panel) or polyclonal anti-INS19 antiserum (right panel). Lane M: protein marker.
    Pcr Analysis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 566 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr analysis/product/Thermo Fisher
    Average 99 stars, based on 566 article reviews
    Price from $9.99 to $1999.99
    pcr analysis - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    Image Search Results


    miR-138 up regulation leads to hTERT suppression in NB4 cells. A: total RNAs from NB4 cells that were transduced with GFP hsa-miR-138-expressing lentiviruses were extracted at 96 h after removal of the lentivirus-containing medium. hTERT mRNA expression was measured by qRT-PCR. Values were normalized to GAPDH . (n=3; *P

    Journal: International Journal of Molecular and Cellular Medicine

    Article Title: Overexpression of MiR-138 Inhibits Cell Growth and Induces Caspase-mediated Apoptosis in Acute Promyelocytic Leukemia Cell Line

    doi: 10.22088/IJMCM.BUMS.7.1.24

    Figure Lengend Snippet: miR-138 up regulation leads to hTERT suppression in NB4 cells. A: total RNAs from NB4 cells that were transduced with GFP hsa-miR-138-expressing lentiviruses were extracted at 96 h after removal of the lentivirus-containing medium. hTERT mRNA expression was measured by qRT-PCR. Values were normalized to GAPDH . (n=3; *P

    Article Snippet: The prepared cDNA was subjected to quan-titative reverse-transcriptase polymerase chain reaction (qRT-PCR), using Maxima SYBR Green Master mix (Thermo Scientific, Waltham, Massa-chusetts, USA) in the Rotor Gene 6000 Real Time PCR inst rument (Corbett Research, Hilden, Germany).

    Techniques: Transduction, Expressing, Quantitative RT-PCR

    MARF1 binds the CCR4-NOT deadenylase complex and cyclin A mRNA. a Co-immunoprecipitation using anti-HA antibody-conjugated beads followed by Western blotting. Ovary lysates expressing 3xHA-MARF1 (MAT67Tub-Gal4 → UASP-3xHA-MARF1) and those from w 1118 negative control were tested. b Co-immunoprecipitation using anti-HA antibody-conjugated beads followed by Western blotting. Ovary lysates expressing 3xHA-tagged MARF1 (MAT67Tub-Gal4 → UASP-3xHA-MARF1 full-length or fragments) and those from w 1118 negative control were tested. Black triangles indicate the detected 3xHA-tagged MARF1 proteins. c Co-immunoprecipitation using ovary lysates from w 1118 and anti-Not1 antibody or negative control IgG-bound protein G beads followed by Western blotting. d Fold enrichment of mRNAs relative to a control gapdh mRNA that were co-immunoprecipitated with MARF1 by anti-HA antibody-conjugated beads and were eluted from the beads normalized by w 1118 negative control, determined by RNA-immunoprecipitation followed by qRT-PCR. Mean ± SD ( n = 3). P -value

    Journal: Nature Communications

    Article Title: LOTUS domain protein MARF1 binds CCR4-NOT deadenylase complex to post-transcriptionally regulate gene expression in oocytes

    doi: 10.1038/s41467-018-06404-w

    Figure Lengend Snippet: MARF1 binds the CCR4-NOT deadenylase complex and cyclin A mRNA. a Co-immunoprecipitation using anti-HA antibody-conjugated beads followed by Western blotting. Ovary lysates expressing 3xHA-MARF1 (MAT67Tub-Gal4 → UASP-3xHA-MARF1) and those from w 1118 negative control were tested. b Co-immunoprecipitation using anti-HA antibody-conjugated beads followed by Western blotting. Ovary lysates expressing 3xHA-tagged MARF1 (MAT67Tub-Gal4 → UASP-3xHA-MARF1 full-length or fragments) and those from w 1118 negative control were tested. Black triangles indicate the detected 3xHA-tagged MARF1 proteins. c Co-immunoprecipitation using ovary lysates from w 1118 and anti-Not1 antibody or negative control IgG-bound protein G beads followed by Western blotting. d Fold enrichment of mRNAs relative to a control gapdh mRNA that were co-immunoprecipitated with MARF1 by anti-HA antibody-conjugated beads and were eluted from the beads normalized by w 1118 negative control, determined by RNA-immunoprecipitation followed by qRT-PCR. Mean ± SD ( n = 3). P -value

    Article Snippet: qRT-PCR RNA from oocytes was prepared using miRVana (Thermo Fisher Scientific).

    Techniques: Immunoprecipitation, Western Blot, Expressing, Negative Control, Quantitative RT-PCR

    Tethered MARF1 shortens reporter mRNA poly-A tail and reduces reporter protein level. a GFP-5x BoxB reporter structure, harboring a ubiquitous tubulin promoter, GFP-coding sequence, and a 3′ UTR containing five BoxB hairpins. LambdaN-HA-fused control peptide, MARF1, GW182, and Piwi under a UASP promoter were expressed in germline cells using maternal-alpha-tubulin-Gal4 driver. b Confocal images of GFP signal in stage 14 oocytes. Scale bar is 100 μm. c Western blots using stage 14 oocyte lysates. Black triangles indicate the transgenic lambda-HA-fused proteins. d Quantification of band intensities in c . e ePAT assay measuring GFP reporter mRNA poly-A tail length in stage 14 oocytes. The amplified DNA sizes were analyzed on an agarose gel. f ePAT assay measuring poly-A tail lengths of GFP reporter mRNA and a negative control cdk1 mRNA in stage 14 oocytes. Amplified DNA sizes were analyzed by capillary fragment analysis. g Relative abundance of GFP-5xBoxB mRNA normalized by gapdh mRNA determined by qRT-PCR. Mean ± SD ( n = 6 for the control and n = 4 for all the others)

    Journal: Nature Communications

    Article Title: LOTUS domain protein MARF1 binds CCR4-NOT deadenylase complex to post-transcriptionally regulate gene expression in oocytes

    doi: 10.1038/s41467-018-06404-w

    Figure Lengend Snippet: Tethered MARF1 shortens reporter mRNA poly-A tail and reduces reporter protein level. a GFP-5x BoxB reporter structure, harboring a ubiquitous tubulin promoter, GFP-coding sequence, and a 3′ UTR containing five BoxB hairpins. LambdaN-HA-fused control peptide, MARF1, GW182, and Piwi under a UASP promoter were expressed in germline cells using maternal-alpha-tubulin-Gal4 driver. b Confocal images of GFP signal in stage 14 oocytes. Scale bar is 100 μm. c Western blots using stage 14 oocyte lysates. Black triangles indicate the transgenic lambda-HA-fused proteins. d Quantification of band intensities in c . e ePAT assay measuring GFP reporter mRNA poly-A tail length in stage 14 oocytes. The amplified DNA sizes were analyzed on an agarose gel. f ePAT assay measuring poly-A tail lengths of GFP reporter mRNA and a negative control cdk1 mRNA in stage 14 oocytes. Amplified DNA sizes were analyzed by capillary fragment analysis. g Relative abundance of GFP-5xBoxB mRNA normalized by gapdh mRNA determined by qRT-PCR. Mean ± SD ( n = 6 for the control and n = 4 for all the others)

    Article Snippet: qRT-PCR RNA from oocytes was prepared using miRVana (Thermo Fisher Scientific).

    Techniques: Sequencing, Western Blot, Transgenic Assay, Amplification, Agarose Gel Electrophoresis, Negative Control, Quantitative RT-PCR

    Knockdown or suppression of p65 reduces GLS1 expression in HCC cells. ( A ) Western blot shows the knockdown of c-Myc without interfering with GLS1 expression in HepG2 and Hep3B cells. ( B ) Western blot shows that p65 knockdown reduces GLS1 protein levels in both HepG2 and Hep3B cells. ( C ) qRT-PCR indicates that p65 knockdown reduces GLS1 mRNA levels in both HepG2 and Hep3B cells. ** p

    Journal: OncoTargets and therapy

    Article Title: Nuclear factor-κB p65 regulates glutaminase 1 expression in human hepatocellular carcinoma

    doi: 10.2147/OTT.S167408

    Figure Lengend Snippet: Knockdown or suppression of p65 reduces GLS1 expression in HCC cells. ( A ) Western blot shows the knockdown of c-Myc without interfering with GLS1 expression in HepG2 and Hep3B cells. ( B ) Western blot shows that p65 knockdown reduces GLS1 protein levels in both HepG2 and Hep3B cells. ( C ) qRT-PCR indicates that p65 knockdown reduces GLS1 mRNA levels in both HepG2 and Hep3B cells. ** p

    Article Snippet: Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) Upon relative treatment, cells were lysed, and total RNA was isolated using TRIzol (Thermo Fisher Scientific, Waltham, MA, USA).

    Techniques: Expressing, Western Blot, Quantitative RT-PCR

    Production, purification, and characterization of the INS19 fragment expressed by cgd6_5510 . (A) Electrophoresis analysis of a PCR product from the cgd6_5510 region. Lane M: DNA marker; Lane 1: cgd6_5510 PCR product. (B) The Western blot analysis of the expression of recombinant INS19 in Escherichia coli Rosetta (DE3). Lane M: protein marker; Lane 1: lysate of un-induced bacterial cells transformed with pET28s- cgd6_5510 ; Lane 2: supernatant from lysate of bacterial cells induced by IPTG; Lane 3: inclusion bodies from lysate of bacterial cells induced by IPTG. (C) Purification of recombinant INS19 in inclusion bodies as indicated by results of SDS-PAGE analysis. Lane M: protein marker; Lane 1: proteins from inclusion bodies dissolved in wash buffer containing 1 M urea; Lane 2: proteins from inclusion bodies dissolved in wash buffer containing 2 M urea; Lane 3: proteins from inclusion bodies dissolved in buffer containing 8 M urea; Lane 4: undissolved proteins from inclusion bodies after washing; Lane 5: purified recombinant INS19 after dialysis and ultrafiltration. (D) The Western blot analysis of purified INS19 (Lane 1), C. parvum sporozoite lysate (Lane 2), and C. parvum oocyst lysate (Lane 3) probed with the pre-immune serum (left panel) or polyclonal anti-INS19 antiserum (right panel). Lane M: protein marker.

    Journal: Frontiers in Microbiology

    Article Title: Characterization of a Species-Specific Insulinase-Like Protease in Cryptosporidium parvum

    doi: 10.3389/fmicb.2019.00354

    Figure Lengend Snippet: Production, purification, and characterization of the INS19 fragment expressed by cgd6_5510 . (A) Electrophoresis analysis of a PCR product from the cgd6_5510 region. Lane M: DNA marker; Lane 1: cgd6_5510 PCR product. (B) The Western blot analysis of the expression of recombinant INS19 in Escherichia coli Rosetta (DE3). Lane M: protein marker; Lane 1: lysate of un-induced bacterial cells transformed with pET28s- cgd6_5510 ; Lane 2: supernatant from lysate of bacterial cells induced by IPTG; Lane 3: inclusion bodies from lysate of bacterial cells induced by IPTG. (C) Purification of recombinant INS19 in inclusion bodies as indicated by results of SDS-PAGE analysis. Lane M: protein marker; Lane 1: proteins from inclusion bodies dissolved in wash buffer containing 1 M urea; Lane 2: proteins from inclusion bodies dissolved in wash buffer containing 2 M urea; Lane 3: proteins from inclusion bodies dissolved in buffer containing 8 M urea; Lane 4: undissolved proteins from inclusion bodies after washing; Lane 5: purified recombinant INS19 after dialysis and ultrafiltration. (D) The Western blot analysis of purified INS19 (Lane 1), C. parvum sporozoite lysate (Lane 2), and C. parvum oocyst lysate (Lane 3) probed with the pre-immune serum (left panel) or polyclonal anti-INS19 antiserum (right panel). Lane M: protein marker.

    Article Snippet: The PCR analysis of DNA and cDNA was performed in duplicates with the Phusion High-Fidelity DNA Polymerase (Thermo Fisher Scientific) under the following conditions: denaturation at 95°C for 5 min; 35 cycles of amplifications at 95°C for 45 s, 50°C for 45 s, and 72°C for 60 s; and a final extension at 72°C for 7 min.

    Techniques: Purification, Electrophoresis, Polymerase Chain Reaction, Marker, Western Blot, Expressing, Recombinant, Transformation Assay, SDS Page