topo xl cloning kit  (Thermo Fisher)


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    Name:
    TOPO TA Cloning Kit
    Description:
    TOPO TA Cloning Dual Promoter kits are for fast efficient cloning and subsequent in vitro transcription translation These kits includes the pCR II TOPO TA vector with dual T7 and SP6 promoters By eliminating time consuming and tedious restriction site cloning TOPO cloning is the most reliable cloning method featuring a 5 minute cloning reaction adn 3 step protocol and yielding up to 95 recombinants Convenient features of this TOPO TA Cloning Dual Promoter kit include • 3 T overhangs for direct ligation of Taq amplified PCR products • T7 and SP6 promoters for efficient in vitro transcription • M13 forward and reverse primer sites for sequencing • 16 convenient restriction sites including EcoRI flanking your insert for subsequent excision or subcloning • Kanamycin and ampicillin resistance for your choice of selection in E coli • Easy blue white colony screening for selection of recombinants Kit Options TOPO TA Cloning Dual Promoter Kits TOPO TA Cloning Dual Promoter kits can be purchased with a variety of competent cells that deliver different advantages depending upon your needs • General cloning TOP10 cells Cat Nos K4600 J10 K4600 01 K4600 40 • High efficiency cloning TOP10 Electrocomp cells Cat Nos K4660 01 K4660 40 • General cloning bacteriophage T1 resistance DH5α T1R cells Cat Nos K4620 01 K4620 40 • Fast growth Mach1 T1R chemically competent E coli Cat No K4610 20 • Repressor induction needs TOP10F cells Cat Nos K4650 01 K4650 40 • Provide your own cells Cat Nos 450640 and 452640
    Catalog Number:
    450640
    Price:
    None
    Applications:
    Cloning|PCR Cloning
    Category:
    DNA Vectors Clones Purified Nucleic Acids Libraries
    Buy from Supplier


    Structured Review

    Thermo Fisher topo xl cloning kit
    Summary of the strategy adopted to obtain the M. persicae RyR cDNA sequence. Five overlapping fragments (F1a/b, F2, F3 and <t>F4)</t> were initially PCR amplified. Fragments F1a, F1b, F2 and F3 were sub-cloned into pJET1.2blunt vector (Thermo-Fermentas) and fragment F4 was sub-cloned into a <t>TOPO®TA</t> vector (Invitrogen) for sequencing.
    TOPO TA Cloning Dual Promoter kits are for fast efficient cloning and subsequent in vitro transcription translation These kits includes the pCR II TOPO TA vector with dual T7 and SP6 promoters By eliminating time consuming and tedious restriction site cloning TOPO cloning is the most reliable cloning method featuring a 5 minute cloning reaction adn 3 step protocol and yielding up to 95 recombinants Convenient features of this TOPO TA Cloning Dual Promoter kit include • 3 T overhangs for direct ligation of Taq amplified PCR products • T7 and SP6 promoters for efficient in vitro transcription • M13 forward and reverse primer sites for sequencing • 16 convenient restriction sites including EcoRI flanking your insert for subsequent excision or subcloning • Kanamycin and ampicillin resistance for your choice of selection in E coli • Easy blue white colony screening for selection of recombinants Kit Options TOPO TA Cloning Dual Promoter Kits TOPO TA Cloning Dual Promoter kits can be purchased with a variety of competent cells that deliver different advantages depending upon your needs • General cloning TOP10 cells Cat Nos K4600 J10 K4600 01 K4600 40 • High efficiency cloning TOP10 Electrocomp cells Cat Nos K4660 01 K4660 40 • General cloning bacteriophage T1 resistance DH5α T1R cells Cat Nos K4620 01 K4620 40 • Fast growth Mach1 T1R chemically competent E coli Cat No K4610 20 • Repressor induction needs TOP10F cells Cat Nos K4650 01 K4650 40 • Provide your own cells Cat Nos 450640 and 452640
    https://www.bioz.com/result/topo xl cloning kit/product/Thermo Fisher
    Average 90 stars, based on 25 article reviews
    Price from $9.99 to $1999.99
    topo xl cloning kit - by Bioz Stars, 2020-02
    90/100 stars

    Images

    1) Product Images from "Molecular cloning, characterisation and mRNA expression of the ryanodine receptor from the peach-potato aphid, Myzus persicae"

    Article Title: Molecular cloning, characterisation and mRNA expression of the ryanodine receptor from the peach-potato aphid, Myzus persicae

    Journal: Gene

    doi: 10.1016/j.gene.2014.11.035

    Summary of the strategy adopted to obtain the M. persicae RyR cDNA sequence. Five overlapping fragments (F1a/b, F2, F3 and F4) were initially PCR amplified. Fragments F1a, F1b, F2 and F3 were sub-cloned into pJET1.2blunt vector (Thermo-Fermentas) and fragment F4 was sub-cloned into a TOPO®TA vector (Invitrogen) for sequencing.
    Figure Legend Snippet: Summary of the strategy adopted to obtain the M. persicae RyR cDNA sequence. Five overlapping fragments (F1a/b, F2, F3 and F4) were initially PCR amplified. Fragments F1a, F1b, F2 and F3 were sub-cloned into pJET1.2blunt vector (Thermo-Fermentas) and fragment F4 was sub-cloned into a TOPO®TA vector (Invitrogen) for sequencing.

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

    Related Articles

    Methylation Sequencing:

    Article Title: Epigenetic silencing of genes and microRNAs within the imprinted Dlk1-Dio3 region at human chromosome 14.32 in giant cell tumor of bone
    Article Snippet: Paragraph title: Bisulfite sequencing ... PCR products were cloned into pCR4-TOPO vector using TOPO TA cloning kit (Life Technologies) and sequenced.

    Article Title: Epigenetic and Genetic Alterations in Netrin-1 Receptors UNC5C and DCC in Human Colon Cancer
    Article Snippet: Paragraph title: Cloning and Sodium Bisulfite Sequencing ... Each of the PCR products subsequently was cloned using the TOPO-TA cloning system (Invitrogen Life Technologies Inc).

    Clone Assay:

    Article Title: Genetic transformation of novel isolates of chicken Lactobacillus bearing probiotic features for expression of heterologous proteins: a tool to develop live oral vaccines
    Article Snippet: .. PCR products were cleaned-up using the Concert™ Rapid PCR Purification System and cloned in E. coli XL1-Blue into pCR2.1-TOPO vector using the Invitrogen TOPO TA cloning kit (Invitrogen Life Technologies, Carlsbad, USA). .. PCR was performed on cell lysates of ampicillin-resistant transformants using M13 specific primers to confirm the size of the inserts.

    Article Title: In vivo dissection of the chromosome condensation machinery
    Article Snippet: .. PCR cloning was performed using the TOPO-TA cloning kit (Invitrogen). rDNA and CEN16 proximal FISH probes were generated as previously described ( ). .. Antibodies used were as follows: mAb YOL1/34 (rat anti-tubulin; Serotec), mAb 12CA5 (anti-HA; BAbCo), mouse antidigoxigenin and pig anti–goat-FITC (Boehringer), goat anti–mouse-FITC (BAbCo), rabbit anti-GFP (CLONTECH Laboratories, Inc.), and goat anti–rat-FITC and CY3 anti–rabbit (Jackson ImmunoResearch Laboratories).

    Article Title: Duplication and concerted evolution of MiSp-encoding genes underlie the material properties of minor ampullate silks of cobweb weaving spiders
    Article Snippet: .. Cycling conditions were 40 cycles of 98 ° C for 5 s, 58 ° C for 15 s, and 72 ° C for 2.5 min. PCR products were gel-excised and cloned using the TOPO®-Blunt Cloning® Kit (Invitrogen). .. End sequencing of these clones revealed numerous premature stop codons and thus clones were not completely sequenced.

    Article Title: Characterization of HCV Interactions with Toll-Like Receptors and RIG-I in Liver Cells
    Article Snippet: .. Core, Envelope E1/E2 and NS3/4A were amplified from pJFH-1 plasmid and cloned using TOPO TA Cloning kit from Invitrogen ( shows PCR primers used for amplification). .. Cells were transfected using Lipofectamine following the manufacturer's recommendations (Invitrogen).

    Article Title: Epigenetic silencing of genes and microRNAs within the imprinted Dlk1-Dio3 region at human chromosome 14.32 in giant cell tumor of bone
    Article Snippet: .. PCR products were cloned into pCR4-TOPO vector using TOPO TA cloning kit (Life Technologies) and sequenced. .. Methylation was analyzed using BiQ-Analyzer software [ ].

    Article Title: Strategy for efficient generation of numerous full-length cDNA clones of classical swine fever virus for haplotyping
    Article Snippet: .. The cDNA amplicons were also cloned using the TOPO XL-2 kit (Thermo Scientific). .. Briefly, the linearized pCR-XL-2-TOPO vector was mixed with amplicons (ca.

    Article Title: Duplication and concerted evolution of MiSp-encoding genes underlie the material properties of minor ampullate silks of cobweb weaving spiders
    Article Snippet: .. Cycling conditions were 40 cycles of 94 ° C for 30 s, 50–60 ° C (depending on primer pair) for 45 s, and 68 ° C for 10 min. PCR products were gel-excised and cloned using the TOPO®-TA Cloning® Kit (Invitrogen). .. Clones were screened by PCR amplification of MiSp N and/or C-terminal encoding region, and MiSp positive clones were sequenced with Sp6, T7, M13F, or M13R universal primers.

    Article Title: Generation of TALEN-Mediated GRdim Knock-In Rats by Homologous Recombination
    Article Snippet: .. Amplified DNA was cloned into blunt vector using the TOPO cloning kit (Invitrogen, now Life Technologies) and transformed into HB101 bacteria. ..

    Article Title: Calcium Influx and DREAM Protein Are Required For GnRH Gene Expression Pulse Activity
    Article Snippet: .. The amplified RT-PCR fragment DAM12 (852 bp) was cloned into pCR2.1-TOPO vector using the TOPO TA Cloning Kit (Invitrogen). .. The resulting plasmids were purified using the QIAprep Spin Miniprep Kits (Qiagen Inc., Valencia, CA).

    Article Title: CRISPR/Cas9‐mediated knockout of six glycosyltransferase genes in Nicotiana benthamiana for the production of recombinant proteins lacking β‐1,2‐xylose and core α‐1,3‐fucose
    Article Snippet: .. When the sequencing results were ambiguous, the PCR products were cloned in pTOPO from the TOPO TA Cloning Kit (Thermo Fisher Scientific, Waltham) and 5–10 clones were sequenced individually for each sample. ..

    Article Title: Rapid generation of gene disruption constructs by in vitro transposition and identification of a Dictyostelium protein kinase that regulates its rate of growth and development
    Article Snippet: .. The target gene was PCR amplified using the genomic DNA as template and, after gel purification, the amplicons were cloned into the vector pCR2.1-TOPO by topoisomerase cloning (TOPO-TA cloning kit; Invitrogen, USA). .. The transposable bsr cassette was excised from the EZTN:tetr -bsr vector (Fig. ) by PvuII digestion and gel purified.

    Article Title: Inactivation of class II transactivator by DNA methylation and histone deacetylation associated with absence of HLA-DR induction by interferon-γ in haematopoietic tumour cells
    Article Snippet: .. For bisulphite sequencing, the PCR products were amplified using primers CIITA-GM1F and CIITA-SEQR, 5′-ACAATCTCRAAACCTCRATTCTC-3′, and cloned into pCR4 vector using a TOPO-TA cloning Kit (Invitrogen), after which the plasmid DNA was purified using a PI system (Kurabo, Tokyo, Japan). .. Sequencing was carried out using a BigDye Terminator Kit and an ABI 3100 DNA sequencer (Applied Biosystems).

    Article Title: Identification of Differentially Expressed Genes in Papillary Thyroid Cancers
    Article Snippet: .. Detection of gene from cloning and sequencing The mRNAs of the DEGs showing distinct results were extracted, amplified using the TOPO TA Cloning kit (Invitrogen, K4500-01, CA, USA) and were sequenced. .. The DNA sequence of each gene was confirmed by comparison with sequences in GenBank (NIH, MD, USA). shows 10 DEGs accessed by GeneBank and their homologies.

    Article Title: Epigenetic and Genetic Alterations in Netrin-1 Receptors UNC5C and DCC in Human Colon Cancer
    Article Snippet: .. Each of the PCR products subsequently was cloned using the TOPO-TA cloning system (Invitrogen Life Technologies Inc). .. Nine to 11 white colonies indicating positive clones with PCR products were sequenced using ABI PRISM Big Dye Terminator v1.1 Cycle Sequencing Kits on an ABI PRISM 3100 Avant Genetic analyzer (Applied Biosystems, Foster City, CA).

    Amplification:

    Article Title: Duplication and concerted evolution of MiSp-encoding genes underlie the material properties of minor ampullate silks of cobweb weaving spiders
    Article Snippet: Paragraph title: Amplification and cloning of S. grossa and P. tepidariorum MiSp ... Cycling conditions were 40 cycles of 98 ° C for 5 s, 58 ° C for 15 s, and 72 ° C for 2.5 min. PCR products were gel-excised and cloned using the TOPO®-Blunt Cloning® Kit (Invitrogen).

    Article Title: Characterization of HCV Interactions with Toll-Like Receptors and RIG-I in Liver Cells
    Article Snippet: .. Core, Envelope E1/E2 and NS3/4A were amplified from pJFH-1 plasmid and cloned using TOPO TA Cloning kit from Invitrogen ( shows PCR primers used for amplification). .. Cells were transfected using Lipofectamine following the manufacturer's recommendations (Invitrogen).

    Article Title: Epigenetic silencing of genes and microRNAs within the imprinted Dlk1-Dio3 region at human chromosome 14.32 in giant cell tumor of bone
    Article Snippet: DNA fragments covering the IG-DMR and the Meg3-DMR, respectively, were amplified by PCR using the following primers: IG-DMR-F: 5′-TGGGATTATAGGTATTATGTTTGGA-3′, IG-DMR-R: 5′-CACTACTAAAAACTACATTTAAACAA-3′, Meg3DMR-F 5′- GTTAGGGATTAATTTTTATGTGTTAG-3′, Meg3DMR-R 5′-CAAATTCTATAACAAATTACTCTAAC-3′. .. PCR products were cloned into pCR4-TOPO vector using TOPO TA cloning kit (Life Technologies) and sequenced.

    Article Title: Generation of TALEN-Mediated GRdim Knock-In Rats by Homologous Recombination
    Article Snippet: .. Amplified DNA was cloned into blunt vector using the TOPO cloning kit (Invitrogen, now Life Technologies) and transformed into HB101 bacteria. ..

    Article Title: Calcium Influx and DREAM Protein Are Required For GnRH Gene Expression Pulse Activity
    Article Snippet: .. The amplified RT-PCR fragment DAM12 (852 bp) was cloned into pCR2.1-TOPO vector using the TOPO TA Cloning Kit (Invitrogen). .. The resulting plasmids were purified using the QIAprep Spin Miniprep Kits (Qiagen Inc., Valencia, CA).

    Article Title: CRISPR/Cas9‐mediated knockout of six glycosyltransferase genes in Nicotiana benthamiana for the production of recombinant proteins lacking β‐1,2‐xylose and core α‐1,3‐fucose
    Article Snippet: Paragraph title: Amplification of target regions and Sanger sequencing of amplicons ... When the sequencing results were ambiguous, the PCR products were cloned in pTOPO from the TOPO TA Cloning Kit (Thermo Fisher Scientific, Waltham) and 5–10 clones were sequenced individually for each sample.

    Article Title: Rapid generation of gene disruption constructs by in vitro transposition and identification of a Dictyostelium protein kinase that regulates its rate of growth and development
    Article Snippet: .. The target gene was PCR amplified using the genomic DNA as template and, after gel purification, the amplicons were cloned into the vector pCR2.1-TOPO by topoisomerase cloning (TOPO-TA cloning kit; Invitrogen, USA). .. The transposable bsr cassette was excised from the EZTN:tetr -bsr vector (Fig. ) by PvuII digestion and gel purified.

    Article Title: Inactivation of class II transactivator by DNA methylation and histone deacetylation associated with absence of HLA-DR induction by interferon-γ in haematopoietic tumour cells
    Article Snippet: .. For bisulphite sequencing, the PCR products were amplified using primers CIITA-GM1F and CIITA-SEQR, 5′-ACAATCTCRAAACCTCRATTCTC-3′, and cloned into pCR4 vector using a TOPO-TA cloning Kit (Invitrogen), after which the plasmid DNA was purified using a PI system (Kurabo, Tokyo, Japan). .. Sequencing was carried out using a BigDye Terminator Kit and an ABI 3100 DNA sequencer (Applied Biosystems).

    Article Title: Identification of Differentially Expressed Genes in Papillary Thyroid Cancers
    Article Snippet: .. Detection of gene from cloning and sequencing The mRNAs of the DEGs showing distinct results were extracted, amplified using the TOPO TA Cloning kit (Invitrogen, K4500-01, CA, USA) and were sequenced. .. The DNA sequence of each gene was confirmed by comparison with sequences in GenBank (NIH, MD, USA). shows 10 DEGs accessed by GeneBank and their homologies.

    Article Title: Epigenetic and Genetic Alterations in Netrin-1 Receptors UNC5C and DCC in Human Colon Cancer
    Article Snippet: Bisulfite-modified DNA from cell lines and clinical specimens was amplified using primer sets Bis-F and COBRA-R ( ). .. Each of the PCR products subsequently was cloned using the TOPO-TA cloning system (Invitrogen Life Technologies Inc).

    TA Cloning:

    Article Title: Genetic transformation of novel isolates of chicken Lactobacillus bearing probiotic features for expression of heterologous proteins: a tool to develop live oral vaccines
    Article Snippet: .. PCR products were cleaned-up using the Concert™ Rapid PCR Purification System and cloned in E. coli XL1-Blue into pCR2.1-TOPO vector using the Invitrogen TOPO TA cloning kit (Invitrogen Life Technologies, Carlsbad, USA). .. PCR was performed on cell lysates of ampicillin-resistant transformants using M13 specific primers to confirm the size of the inserts.

    Article Title: Characterization of HCV Interactions with Toll-Like Receptors and RIG-I in Liver Cells
    Article Snippet: .. Core, Envelope E1/E2 and NS3/4A were amplified from pJFH-1 plasmid and cloned using TOPO TA Cloning kit from Invitrogen ( shows PCR primers used for amplification). .. Cells were transfected using Lipofectamine following the manufacturer's recommendations (Invitrogen).

    Article Title: Epigenetic silencing of genes and microRNAs within the imprinted Dlk1-Dio3 region at human chromosome 14.32 in giant cell tumor of bone
    Article Snippet: .. PCR products were cloned into pCR4-TOPO vector using TOPO TA cloning kit (Life Technologies) and sequenced. .. Methylation was analyzed using BiQ-Analyzer software [ ].

    Article Title: Calcium Influx and DREAM Protein Are Required For GnRH Gene Expression Pulse Activity
    Article Snippet: .. The amplified RT-PCR fragment DAM12 (852 bp) was cloned into pCR2.1-TOPO vector using the TOPO TA Cloning Kit (Invitrogen). .. The resulting plasmids were purified using the QIAprep Spin Miniprep Kits (Qiagen Inc., Valencia, CA).

    Article Title: Single-cell analysis of the fate of c-kit-positive bone marrow cells
    Article Snippet: .. Taq polymerase-amplified PCR products were inserted into the plasmid vector pCR4-TOPO using the TOPO TA Cloning Kit (Invitrogen). .. Subsequently, chemically competent TOP10 E. coli cells were transformed with the vector carrying the PCR products.

    Article Title: CRISPR/Cas9‐mediated knockout of six glycosyltransferase genes in Nicotiana benthamiana for the production of recombinant proteins lacking β‐1,2‐xylose and core α‐1,3‐fucose
    Article Snippet: .. When the sequencing results were ambiguous, the PCR products were cloned in pTOPO from the TOPO TA Cloning Kit (Thermo Fisher Scientific, Waltham) and 5–10 clones were sequenced individually for each sample. ..

    Article Title: Identification of Differentially Expressed Genes in Papillary Thyroid Cancers
    Article Snippet: .. Detection of gene from cloning and sequencing The mRNAs of the DEGs showing distinct results were extracted, amplified using the TOPO TA Cloning kit (Invitrogen, K4500-01, CA, USA) and were sequenced. .. The DNA sequence of each gene was confirmed by comparison with sequences in GenBank (NIH, MD, USA). shows 10 DEGs accessed by GeneBank and their homologies.

    Incubation:

    Article Title: Characterization of HCV Interactions with Toll-Like Receptors and RIG-I in Liver Cells
    Article Snippet: Core, Envelope E1/E2 and NS3/4A were amplified from pJFH-1 plasmid and cloned using TOPO TA Cloning kit from Invitrogen ( shows PCR primers used for amplification). .. Briefly, in a 6-well tissue culture plate (Fisherbrand), 1×105 Huh7.5 or LH86 cells were seeded in 2 mL of cDMEM and incubated at 37°C overnight.

    Article Title: Strategy for efficient generation of numerous full-length cDNA clones of classical swine fever virus for haplotyping
    Article Snippet: The cDNA amplicons were also cloned using the TOPO XL-2 kit (Thermo Scientific). .. The mixtures of vector and amplicons were incubated at 25 °C for 30 min. A 2-μl aliquot was used to transform E. coli One Shot™ OmniMAX™ 2TIR competent cells according to the manufacturer’s protocol, and colonies were grown on LB plates containing kanamycin (KAN) (50 μg/ml) and 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG).

    Article Title: Generation of TALEN-Mediated GRdim Knock-In Rats by Homologous Recombination
    Article Snippet: T7 Endo I mismatch detection assay and cloning 1 µl of NEB Buffer 2 (New England Biolabs) and, 0.5 µl of T7 Endo I nuclease (New England Biolabs) were added to the PCR mix previously prepared and incubated for 30 minutes at 37°C. .. Amplified DNA was cloned into blunt vector using the TOPO cloning kit (Invitrogen, now Life Technologies) and transformed into HB101 bacteria.

    Article Title: Single-cell analysis of the fate of c-kit-positive bone marrow cells
    Article Snippet: Taq polymerase-amplified PCR products were inserted into the plasmid vector pCR4-TOPO using the TOPO TA Cloning Kit (Invitrogen). .. The transformation mixture was spread on agar plates and incubated overnight at 37 °C.

    Expressing:

    Article Title: Characterization of HCV Interactions with Toll-Like Receptors and RIG-I in Liver Cells
    Article Snippet: The expression vector pTOPO was from Invitrogen (Carlsbad, CA) and the negative control siRNA was produced by T7 polymerase in vitro with the Ambion Silencer siRNA construction Kit or purchased (both control and kit from Ambion, Austin, TX). .. Core, Envelope E1/E2 and NS3/4A were amplified from pJFH-1 plasmid and cloned using TOPO TA Cloning kit from Invitrogen ( shows PCR primers used for amplification).

    Transformation Assay:

    Article Title: Generation of TALEN-Mediated GRdim Knock-In Rats by Homologous Recombination
    Article Snippet: .. Amplified DNA was cloned into blunt vector using the TOPO cloning kit (Invitrogen, now Life Technologies) and transformed into HB101 bacteria. ..

    Article Title: Single-cell analysis of the fate of c-kit-positive bone marrow cells
    Article Snippet: Taq polymerase-amplified PCR products were inserted into the plasmid vector pCR4-TOPO using the TOPO TA Cloning Kit (Invitrogen). .. Subsequently, chemically competent TOP10 E. coli cells were transformed with the vector carrying the PCR products.

    Bisulfite Sequencing:

    Article Title: Inactivation of class II transactivator by DNA methylation and histone deacetylation associated with absence of HLA-DR induction by interferon-γ in haematopoietic tumour cells
    Article Snippet: .. For bisulphite sequencing, the PCR products were amplified using primers CIITA-GM1F and CIITA-SEQR, 5′-ACAATCTCRAAACCTCRATTCTC-3′, and cloned into pCR4 vector using a TOPO-TA cloning Kit (Invitrogen), after which the plasmid DNA was purified using a PI system (Kurabo, Tokyo, Japan). .. Sequencing was carried out using a BigDye Terminator Kit and an ABI 3100 DNA sequencer (Applied Biosystems).

    Transfection:

    Article Title: Characterization of HCV Interactions with Toll-Like Receptors and RIG-I in Liver Cells
    Article Snippet: Core, Envelope E1/E2 and NS3/4A were amplified from pJFH-1 plasmid and cloned using TOPO TA Cloning kit from Invitrogen ( shows PCR primers used for amplification). .. Cells were transfected using Lipofectamine following the manufacturer's recommendations (Invitrogen).

    Sequencing:

    Article Title: Duplication and concerted evolution of MiSp-encoding genes underlie the material properties of minor ampullate silks of cobweb weaving spiders
    Article Snippet: Cycling conditions were 40 cycles of 98 ° C for 5 s, 58 ° C for 15 s, and 72 ° C for 2.5 min. PCR products were gel-excised and cloned using the TOPO®-Blunt Cloning® Kit (Invitrogen). .. End sequencing of these clones revealed numerous premature stop codons and thus clones were not completely sequenced.

    Article Title: Epigenetic silencing of genes and microRNAs within the imprinted Dlk1-Dio3 region at human chromosome 14.32 in giant cell tumor of bone
    Article Snippet: According to the sequence NT_026437.12 at NCBI Database the position of the analyzed IG-DMR sequence is 82.276.640 – 82.277.549 and that of the analyzed Meg3-DMR fragment is 82.291.515 – 82.292.333. .. PCR products were cloned into pCR4-TOPO vector using TOPO TA cloning kit (Life Technologies) and sequenced.

    Article Title: Duplication and concerted evolution of MiSp-encoding genes underlie the material properties of minor ampullate silks of cobweb weaving spiders
    Article Snippet: Cycling conditions were 40 cycles of 94 ° C for 30 s, 50–60 ° C (depending on primer pair) for 45 s, and 68 ° C for 10 min. PCR products were gel-excised and cloned using the TOPO®-TA Cloning® Kit (Invitrogen). .. End sequencing of TOPO clones indicated that, within both L. geometricus and L. tredecimguttatus , clones clustered into two distinct groups (Additional file : Figure S2).

    Article Title: Calcium Influx and DREAM Protein Are Required For GnRH Gene Expression Pulse Activity
    Article Snippet: These primers has been used successfully to amplify specifically the whole coding sequence of the rat DREAM gene from the pituitary ( ). .. The amplified RT-PCR fragment DAM12 (852 bp) was cloned into pCR2.1-TOPO vector using the TOPO TA Cloning Kit (Invitrogen).

    Article Title: CRISPR/Cas9‐mediated knockout of six glycosyltransferase genes in Nicotiana benthamiana for the production of recombinant proteins lacking β‐1,2‐xylose and core α‐1,3‐fucose
    Article Snippet: .. When the sequencing results were ambiguous, the PCR products were cloned in pTOPO from the TOPO TA Cloning Kit (Thermo Fisher Scientific, Waltham) and 5–10 clones were sequenced individually for each sample. ..

    Article Title: Inactivation of class II transactivator by DNA methylation and histone deacetylation associated with absence of HLA-DR induction by interferon-γ in haematopoietic tumour cells
    Article Snippet: For bisulphite sequencing, the PCR products were amplified using primers CIITA-GM1F and CIITA-SEQR, 5′-ACAATCTCRAAACCTCRATTCTC-3′, and cloned into pCR4 vector using a TOPO-TA cloning Kit (Invitrogen), after which the plasmid DNA was purified using a PI system (Kurabo, Tokyo, Japan). .. Sequencing was carried out using a BigDye Terminator Kit and an ABI 3100 DNA sequencer (Applied Biosystems).

    Article Title: Identification of Differentially Expressed Genes in Papillary Thyroid Cancers
    Article Snippet: .. Detection of gene from cloning and sequencing The mRNAs of the DEGs showing distinct results were extracted, amplified using the TOPO TA Cloning kit (Invitrogen, K4500-01, CA, USA) and were sequenced. .. The DNA sequence of each gene was confirmed by comparison with sequences in GenBank (NIH, MD, USA). shows 10 DEGs accessed by GeneBank and their homologies.

    Article Title: Epigenetic and Genetic Alterations in Netrin-1 Receptors UNC5C and DCC in Human Colon Cancer
    Article Snippet: Each of the PCR products subsequently was cloned using the TOPO-TA cloning system (Invitrogen Life Technologies Inc). .. Nine to 11 white colonies indicating positive clones with PCR products were sequenced using ABI PRISM Big Dye Terminator v1.1 Cycle Sequencing Kits on an ABI PRISM 3100 Avant Genetic analyzer (Applied Biosystems, Foster City, CA).

    Cell Culture:

    Article Title: Rapid generation of gene disruption constructs by in vitro transposition and identification of a Dictyostelium protein kinase that regulates its rate of growth and development
    Article Snippet: Dictyostelium Ax-2 cells were cultured axenically in HL-5 medium ( ). .. The target gene was PCR amplified using the genomic DNA as template and, after gel purification, the amplicons were cloned into the vector pCR2.1-TOPO by topoisomerase cloning (TOPO-TA cloning kit; Invitrogen, USA).

    Generated:

    Article Title: In vivo dissection of the chromosome condensation machinery
    Article Snippet: .. PCR cloning was performed using the TOPO-TA cloning kit (Invitrogen). rDNA and CEN16 proximal FISH probes were generated as previously described ( ). .. Antibodies used were as follows: mAb YOL1/34 (rat anti-tubulin; Serotec), mAb 12CA5 (anti-HA; BAbCo), mouse antidigoxigenin and pig anti–goat-FITC (Boehringer), goat anti–mouse-FITC (BAbCo), rabbit anti-GFP (CLONTECH Laboratories, Inc.), and goat anti–rat-FITC and CY3 anti–rabbit (Jackson ImmunoResearch Laboratories).

    DNA Sequencing:

    Article Title: Calcium Influx and DREAM Protein Are Required For GnRH Gene Expression Pulse Activity
    Article Snippet: The amplified RT-PCR fragment DAM12 (852 bp) was cloned into pCR2.1-TOPO vector using the TOPO TA Cloning Kit (Invitrogen). .. Integrity and identity of six independent cloned RT-PCR DNA fragments were confirmed by nucleotide DNA sequencing which was performed by the Biotechnology Resource Laboratory of the Medical University of South Carolina (Charleston, SC).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Calcium Influx and DREAM Protein Are Required For GnRH Gene Expression Pulse Activity
    Article Snippet: .. The amplified RT-PCR fragment DAM12 (852 bp) was cloned into pCR2.1-TOPO vector using the TOPO TA Cloning Kit (Invitrogen). .. The resulting plasmids were purified using the QIAprep Spin Miniprep Kits (Qiagen Inc., Valencia, CA).

    Gel Purification:

    Article Title: Rapid generation of gene disruption constructs by in vitro transposition and identification of a Dictyostelium protein kinase that regulates its rate of growth and development
    Article Snippet: .. The target gene was PCR amplified using the genomic DNA as template and, after gel purification, the amplicons were cloned into the vector pCR2.1-TOPO by topoisomerase cloning (TOPO-TA cloning kit; Invitrogen, USA). .. The transposable bsr cassette was excised from the EZTN:tetr -bsr vector (Fig. ) by PvuII digestion and gel purified.

    Cellular Antioxidant Activity Assay:

    Article Title: Calcium Influx and DREAM Protein Are Required For GnRH Gene Expression Pulse Activity
    Article Snippet: For PCR amplification of the mouse DREAM gene, we used the PCR primers DAMF1 (5′-GGG AAG ATT AGT GAC GG) and DAMR2 (5′-CTC TGG GTT GAG AAG CAA TGA) which were based on the mouse DREAM cDNA sequence (GeneBank Accession Number: ) and predicted to amplify a fragment of 852 bp. .. The amplified RT-PCR fragment DAM12 (852 bp) was cloned into pCR2.1-TOPO vector using the TOPO TA Cloning Kit (Invitrogen).

    Combined Bisulfite Restriction Analysis Assay:

    Article Title: Inactivation of class II transactivator by DNA methylation and histone deacetylation associated with absence of HLA-DR induction by interferon-γ in haematopoietic tumour cells
    Article Snippet: Paragraph title: Combined bisulphite restriction analysis (COBRA) and bisulphite sequencing ... For bisulphite sequencing, the PCR products were amplified using primers CIITA-GM1F and CIITA-SEQR, 5′-ACAATCTCRAAACCTCRATTCTC-3′, and cloned into pCR4 vector using a TOPO-TA cloning Kit (Invitrogen), after which the plasmid DNA was purified using a PI system (Kurabo, Tokyo, Japan).

    Article Title: Epigenetic and Genetic Alterations in Netrin-1 Receptors UNC5C and DCC in Human Colon Cancer
    Article Snippet: Bisulfite-modified DNA from cell lines and clinical specimens was amplified using primer sets Bis-F and COBRA-R ( ). .. Each of the PCR products subsequently was cloned using the TOPO-TA cloning system (Invitrogen Life Technologies Inc).

    Methylation:

    Article Title: Epigenetic silencing of genes and microRNAs within the imprinted Dlk1-Dio3 region at human chromosome 14.32 in giant cell tumor of bone
    Article Snippet: Bisulfite sequencing For methylation analysis of the IG-DMR and the Meg3-DMR, total cellular DNA was extracted using DNeasy Tissue kit (Qiagen) according to manufacturer’s protocol. .. PCR products were cloned into pCR4-TOPO vector using TOPO TA cloning kit (Life Technologies) and sequenced.

    Article Title: Inactivation of class II transactivator by DNA methylation and histone deacetylation associated with absence of HLA-DR induction by interferon-γ in haematopoietic tumour cells
    Article Snippet: The PCR products were digested with restriction enzymes that cleave CpG sites retained because of methylation. .. For bisulphite sequencing, the PCR products were amplified using primers CIITA-GM1F and CIITA-SEQR, 5′-ACAATCTCRAAACCTCRATTCTC-3′, and cloned into pCR4 vector using a TOPO-TA cloning Kit (Invitrogen), after which the plasmid DNA was purified using a PI system (Kurabo, Tokyo, Japan).

    Isolation:

    Article Title: Rapid generation of gene disruption constructs by in vitro transposition and identification of a Dictyostelium protein kinase that regulates its rate of growth and development
    Article Snippet: Then 5 × 107 cells, at a density of 5 × 106 cells/ml, were harvested and genomic DNA was isolated from the cells by phenol/chloroform extraction. .. The target gene was PCR amplified using the genomic DNA as template and, after gel purification, the amplicons were cloned into the vector pCR2.1-TOPO by topoisomerase cloning (TOPO-TA cloning kit; Invitrogen, USA).

    Detection Assay:

    Article Title: Generation of TALEN-Mediated GRdim Knock-In Rats by Homologous Recombination
    Article Snippet: Paragraph title: T7 Endo I mismatch detection assay and cloning ... Amplified DNA was cloned into blunt vector using the TOPO cloning kit (Invitrogen, now Life Technologies) and transformed into HB101 bacteria.

    Negative Control:

    Article Title: Characterization of HCV Interactions with Toll-Like Receptors and RIG-I in Liver Cells
    Article Snippet: The expression vector pTOPO was from Invitrogen (Carlsbad, CA) and the negative control siRNA was produced by T7 polymerase in vitro with the Ambion Silencer siRNA construction Kit or purchased (both control and kit from Ambion, Austin, TX). .. Core, Envelope E1/E2 and NS3/4A were amplified from pJFH-1 plasmid and cloned using TOPO TA Cloning kit from Invitrogen ( shows PCR primers used for amplification).

    Purification:

    Article Title: Genetic transformation of novel isolates of chicken Lactobacillus bearing probiotic features for expression of heterologous proteins: a tool to develop live oral vaccines
    Article Snippet: .. PCR products were cleaned-up using the Concert™ Rapid PCR Purification System and cloned in E. coli XL1-Blue into pCR2.1-TOPO vector using the Invitrogen TOPO TA cloning kit (Invitrogen Life Technologies, Carlsbad, USA). .. PCR was performed on cell lysates of ampicillin-resistant transformants using M13 specific primers to confirm the size of the inserts.

    Article Title: Calcium Influx and DREAM Protein Are Required For GnRH Gene Expression Pulse Activity
    Article Snippet: The amplified RT-PCR fragment DAM12 (852 bp) was cloned into pCR2.1-TOPO vector using the TOPO TA Cloning Kit (Invitrogen). .. The resulting plasmids were purified using the QIAprep Spin Miniprep Kits (Qiagen Inc., Valencia, CA).

    Article Title: CRISPR/Cas9‐mediated knockout of six glycosyltransferase genes in Nicotiana benthamiana for the production of recombinant proteins lacking β‐1,2‐xylose and core α‐1,3‐fucose
    Article Snippet: Amplification of target regions and Sanger sequencing of amplicons The gRNA target regions were amplified from 1 μL of genomic DNA extract using Q5 Hot Start High‐Fidelity 2x Master Mix (New England Biolabs) in a 20‐μL reaction, and the products purified either directly or from agarose gels using the NucleoSpin Gel and PCR Clean‐up kit, or by robot‐assisted PCR clean‐up with NucleoFast 96 PCR plates (Macherey‐Nagel). .. When the sequencing results were ambiguous, the PCR products were cloned in pTOPO from the TOPO TA Cloning Kit (Thermo Fisher Scientific, Waltham) and 5–10 clones were sequenced individually for each sample.

    Article Title: Inactivation of class II transactivator by DNA methylation and histone deacetylation associated with absence of HLA-DR induction by interferon-γ in haematopoietic tumour cells
    Article Snippet: .. For bisulphite sequencing, the PCR products were amplified using primers CIITA-GM1F and CIITA-SEQR, 5′-ACAATCTCRAAACCTCRATTCTC-3′, and cloned into pCR4 vector using a TOPO-TA cloning Kit (Invitrogen), after which the plasmid DNA was purified using a PI system (Kurabo, Tokyo, Japan). .. Sequencing was carried out using a BigDye Terminator Kit and an ABI 3100 DNA sequencer (Applied Biosystems).

    Polymerase Chain Reaction:

    Article Title: Genetic transformation of novel isolates of chicken Lactobacillus bearing probiotic features for expression of heterologous proteins: a tool to develop live oral vaccines
    Article Snippet: .. PCR products were cleaned-up using the Concert™ Rapid PCR Purification System and cloned in E. coli XL1-Blue into pCR2.1-TOPO vector using the Invitrogen TOPO TA cloning kit (Invitrogen Life Technologies, Carlsbad, USA). .. PCR was performed on cell lysates of ampicillin-resistant transformants using M13 specific primers to confirm the size of the inserts.

    Article Title: In vivo dissection of the chromosome condensation machinery
    Article Snippet: .. PCR cloning was performed using the TOPO-TA cloning kit (Invitrogen). rDNA and CEN16 proximal FISH probes were generated as previously described ( ). .. Antibodies used were as follows: mAb YOL1/34 (rat anti-tubulin; Serotec), mAb 12CA5 (anti-HA; BAbCo), mouse antidigoxigenin and pig anti–goat-FITC (Boehringer), goat anti–mouse-FITC (BAbCo), rabbit anti-GFP (CLONTECH Laboratories, Inc.), and goat anti–rat-FITC and CY3 anti–rabbit (Jackson ImmunoResearch Laboratories).

    Article Title: Duplication and concerted evolution of MiSp-encoding genes underlie the material properties of minor ampullate silks of cobweb weaving spiders
    Article Snippet: .. Cycling conditions were 40 cycles of 98 ° C for 5 s, 58 ° C for 15 s, and 72 ° C for 2.5 min. PCR products were gel-excised and cloned using the TOPO®-Blunt Cloning® Kit (Invitrogen). .. End sequencing of these clones revealed numerous premature stop codons and thus clones were not completely sequenced.

    Article Title: Characterization of HCV Interactions with Toll-Like Receptors and RIG-I in Liver Cells
    Article Snippet: .. Core, Envelope E1/E2 and NS3/4A were amplified from pJFH-1 plasmid and cloned using TOPO TA Cloning kit from Invitrogen ( shows PCR primers used for amplification). .. Cells were transfected using Lipofectamine following the manufacturer's recommendations (Invitrogen).

    Article Title: Epigenetic silencing of genes and microRNAs within the imprinted Dlk1-Dio3 region at human chromosome 14.32 in giant cell tumor of bone
    Article Snippet: .. PCR products were cloned into pCR4-TOPO vector using TOPO TA cloning kit (Life Technologies) and sequenced. .. Methylation was analyzed using BiQ-Analyzer software [ ].

    Article Title: Strategy for efficient generation of numerous full-length cDNA clones of classical swine fever virus for haplotyping
    Article Snippet: Paragraph title: Cloning of amplicons in pCR XL-2-TOPO vector ... The cDNA amplicons were also cloned using the TOPO XL-2 kit (Thermo Scientific).

    Article Title: Duplication and concerted evolution of MiSp-encoding genes underlie the material properties of minor ampullate silks of cobweb weaving spiders
    Article Snippet: .. Cycling conditions were 40 cycles of 94 ° C for 30 s, 50–60 ° C (depending on primer pair) for 45 s, and 68 ° C for 10 min. PCR products were gel-excised and cloned using the TOPO®-TA Cloning® Kit (Invitrogen). .. Clones were screened by PCR amplification of MiSp N and/or C-terminal encoding region, and MiSp positive clones were sequenced with Sp6, T7, M13F, or M13R universal primers.

    Article Title: Generation of TALEN-Mediated GRdim Knock-In Rats by Homologous Recombination
    Article Snippet: T7 Endo I mismatch detection assay and cloning 1 µl of NEB Buffer 2 (New England Biolabs) and, 0.5 µl of T7 Endo I nuclease (New England Biolabs) were added to the PCR mix previously prepared and incubated for 30 minutes at 37°C. .. Amplified DNA was cloned into blunt vector using the TOPO cloning kit (Invitrogen, now Life Technologies) and transformed into HB101 bacteria.

    Article Title: Calcium Influx and DREAM Protein Are Required For GnRH Gene Expression Pulse Activity
    Article Snippet: The PCR amplified products were separated by 1.0% TAE agarose gel electropheresis and visualized by ethidium bromide staining. .. The amplified RT-PCR fragment DAM12 (852 bp) was cloned into pCR2.1-TOPO vector using the TOPO TA Cloning Kit (Invitrogen).

    Article Title: Single-cell analysis of the fate of c-kit-positive bone marrow cells
    Article Snippet: .. Taq polymerase-amplified PCR products were inserted into the plasmid vector pCR4-TOPO using the TOPO TA Cloning Kit (Invitrogen). .. Subsequently, chemically competent TOP10 E. coli cells were transformed with the vector carrying the PCR products.

    Article Title: CRISPR/Cas9‐mediated knockout of six glycosyltransferase genes in Nicotiana benthamiana for the production of recombinant proteins lacking β‐1,2‐xylose and core α‐1,3‐fucose
    Article Snippet: .. When the sequencing results were ambiguous, the PCR products were cloned in pTOPO from the TOPO TA Cloning Kit (Thermo Fisher Scientific, Waltham) and 5–10 clones were sequenced individually for each sample. ..

    Article Title: Rapid generation of gene disruption constructs by in vitro transposition and identification of a Dictyostelium protein kinase that regulates its rate of growth and development
    Article Snippet: .. The target gene was PCR amplified using the genomic DNA as template and, after gel purification, the amplicons were cloned into the vector pCR2.1-TOPO by topoisomerase cloning (TOPO-TA cloning kit; Invitrogen, USA). .. The transposable bsr cassette was excised from the EZTN:tetr -bsr vector (Fig. ) by PvuII digestion and gel purified.

    Article Title: Inactivation of class II transactivator by DNA methylation and histone deacetylation associated with absence of HLA-DR induction by interferon-γ in haematopoietic tumour cells
    Article Snippet: .. For bisulphite sequencing, the PCR products were amplified using primers CIITA-GM1F and CIITA-SEQR, 5′-ACAATCTCRAAACCTCRATTCTC-3′, and cloned into pCR4 vector using a TOPO-TA cloning Kit (Invitrogen), after which the plasmid DNA was purified using a PI system (Kurabo, Tokyo, Japan). .. Sequencing was carried out using a BigDye Terminator Kit and an ABI 3100 DNA sequencer (Applied Biosystems).

    Article Title: Epigenetic and Genetic Alterations in Netrin-1 Receptors UNC5C and DCC in Human Colon Cancer
    Article Snippet: .. Each of the PCR products subsequently was cloned using the TOPO-TA cloning system (Invitrogen Life Technologies Inc). .. Nine to 11 white colonies indicating positive clones with PCR products were sequenced using ABI PRISM Big Dye Terminator v1.1 Cycle Sequencing Kits on an ABI PRISM 3100 Avant Genetic analyzer (Applied Biosystems, Foster City, CA).

    Gel Extraction:

    Article Title: Genetic transformation of novel isolates of chicken Lactobacillus bearing probiotic features for expression of heterologous proteins: a tool to develop live oral vaccines
    Article Snippet: PCR products were cleaned-up using the Concert™ Rapid PCR Purification System and cloned in E. coli XL1-Blue into pCR2.1-TOPO vector using the Invitrogen TOPO TA cloning kit (Invitrogen Life Technologies, Carlsbad, USA). .. Inserts were digested with restriction enzymes, excised from the gel and purified with the Concert™ Rapid Gel Extraction System according to the manufacturer's instructions (Invitrogen Co.).

    Plasmid Preparation:

    Article Title: Genetic transformation of novel isolates of chicken Lactobacillus bearing probiotic features for expression of heterologous proteins: a tool to develop live oral vaccines
    Article Snippet: .. PCR products were cleaned-up using the Concert™ Rapid PCR Purification System and cloned in E. coli XL1-Blue into pCR2.1-TOPO vector using the Invitrogen TOPO TA cloning kit (Invitrogen Life Technologies, Carlsbad, USA). .. PCR was performed on cell lysates of ampicillin-resistant transformants using M13 specific primers to confirm the size of the inserts.

    Article Title: Characterization of HCV Interactions with Toll-Like Receptors and RIG-I in Liver Cells
    Article Snippet: .. Core, Envelope E1/E2 and NS3/4A were amplified from pJFH-1 plasmid and cloned using TOPO TA Cloning kit from Invitrogen ( shows PCR primers used for amplification). .. Cells were transfected using Lipofectamine following the manufacturer's recommendations (Invitrogen).

    Article Title: Epigenetic silencing of genes and microRNAs within the imprinted Dlk1-Dio3 region at human chromosome 14.32 in giant cell tumor of bone
    Article Snippet: .. PCR products were cloned into pCR4-TOPO vector using TOPO TA cloning kit (Life Technologies) and sequenced. .. Methylation was analyzed using BiQ-Analyzer software [ ].

    Article Title: Strategy for efficient generation of numerous full-length cDNA clones of classical swine fever virus for haplotyping
    Article Snippet: Paragraph title: Cloning of amplicons in pCR XL-2-TOPO vector ... The cDNA amplicons were also cloned using the TOPO XL-2 kit (Thermo Scientific).

    Article Title: Generation of TALEN-Mediated GRdim Knock-In Rats by Homologous Recombination
    Article Snippet: .. Amplified DNA was cloned into blunt vector using the TOPO cloning kit (Invitrogen, now Life Technologies) and transformed into HB101 bacteria. ..

    Article Title: Calcium Influx and DREAM Protein Are Required For GnRH Gene Expression Pulse Activity
    Article Snippet: .. The amplified RT-PCR fragment DAM12 (852 bp) was cloned into pCR2.1-TOPO vector using the TOPO TA Cloning Kit (Invitrogen). .. The resulting plasmids were purified using the QIAprep Spin Miniprep Kits (Qiagen Inc., Valencia, CA).

    Article Title: Single-cell analysis of the fate of c-kit-positive bone marrow cells
    Article Snippet: .. Taq polymerase-amplified PCR products were inserted into the plasmid vector pCR4-TOPO using the TOPO TA Cloning Kit (Invitrogen). .. Subsequently, chemically competent TOP10 E. coli cells were transformed with the vector carrying the PCR products.

    Article Title: Rapid generation of gene disruption constructs by in vitro transposition and identification of a Dictyostelium protein kinase that regulates its rate of growth and development
    Article Snippet: .. The target gene was PCR amplified using the genomic DNA as template and, after gel purification, the amplicons were cloned into the vector pCR2.1-TOPO by topoisomerase cloning (TOPO-TA cloning kit; Invitrogen, USA). .. The transposable bsr cassette was excised from the EZTN:tetr -bsr vector (Fig. ) by PvuII digestion and gel purified.

    Article Title: Inactivation of class II transactivator by DNA methylation and histone deacetylation associated with absence of HLA-DR induction by interferon-γ in haematopoietic tumour cells
    Article Snippet: .. For bisulphite sequencing, the PCR products were amplified using primers CIITA-GM1F and CIITA-SEQR, 5′-ACAATCTCRAAACCTCRATTCTC-3′, and cloned into pCR4 vector using a TOPO-TA cloning Kit (Invitrogen), after which the plasmid DNA was purified using a PI system (Kurabo, Tokyo, Japan). .. Sequencing was carried out using a BigDye Terminator Kit and an ABI 3100 DNA sequencer (Applied Biosystems).

    Software:

    Article Title: Epigenetic silencing of genes and microRNAs within the imprinted Dlk1-Dio3 region at human chromosome 14.32 in giant cell tumor of bone
    Article Snippet: PCR products were cloned into pCR4-TOPO vector using TOPO TA cloning kit (Life Technologies) and sequenced. .. Methylation was analyzed using BiQ-Analyzer software [ ].

    Article Title: CRISPR/Cas9‐mediated knockout of six glycosyltransferase genes in Nicotiana benthamiana for the production of recombinant proteins lacking β‐1,2‐xylose and core α‐1,3‐fucose
    Article Snippet: The amplicons were then sequenced in house using the Sanger method, and the results were analysed using Clone Manager v9 Professional (Scientific & Educational Software, Denver) and the online tool TIDE (Brinkman et al ., ). .. When the sequencing results were ambiguous, the PCR products were cloned in pTOPO from the TOPO TA Cloning Kit (Thermo Fisher Scientific, Waltham) and 5–10 clones were sequenced individually for each sample.

    RNA Extraction:

    Article Title: Calcium Influx and DREAM Protein Are Required For GnRH Gene Expression Pulse Activity
    Article Snippet: Paragraph title: 2.3 RNA Extraction and RT-PCR ... The amplified RT-PCR fragment DAM12 (852 bp) was cloned into pCR2.1-TOPO vector using the TOPO TA Cloning Kit (Invitrogen).

    Agarose Gel Electrophoresis:

    Article Title: Calcium Influx and DREAM Protein Are Required For GnRH Gene Expression Pulse Activity
    Article Snippet: The PCR amplified products were separated by 1.0% TAE agarose gel electropheresis and visualized by ethidium bromide staining. .. The amplified RT-PCR fragment DAM12 (852 bp) was cloned into pCR2.1-TOPO vector using the TOPO TA Cloning Kit (Invitrogen).

    Article Title: Inactivation of class II transactivator by DNA methylation and histone deacetylation associated with absence of HLA-DR induction by interferon-γ in haematopoietic tumour cells
    Article Snippet: After ethanol precipitation, the DNA was subjected to 3% agarose gel electrophoresis and stained with ethidium bromide. .. For bisulphite sequencing, the PCR products were amplified using primers CIITA-GM1F and CIITA-SEQR, 5′-ACAATCTCRAAACCTCRATTCTC-3′, and cloned into pCR4 vector using a TOPO-TA cloning Kit (Invitrogen), after which the plasmid DNA was purified using a PI system (Kurabo, Tokyo, Japan).

    In Vitro:

    Article Title: Characterization of HCV Interactions with Toll-Like Receptors and RIG-I in Liver Cells
    Article Snippet: The expression vector pTOPO was from Invitrogen (Carlsbad, CA) and the negative control siRNA was produced by T7 polymerase in vitro with the Ambion Silencer siRNA construction Kit or purchased (both control and kit from Ambion, Austin, TX). .. Core, Envelope E1/E2 and NS3/4A were amplified from pJFH-1 plasmid and cloned using TOPO TA Cloning kit from Invitrogen ( shows PCR primers used for amplification).

    Ethanol Precipitation:

    Article Title: Inactivation of class II transactivator by DNA methylation and histone deacetylation associated with absence of HLA-DR induction by interferon-γ in haematopoietic tumour cells
    Article Snippet: After ethanol precipitation, the DNA was subjected to 3% agarose gel electrophoresis and stained with ethidium bromide. .. For bisulphite sequencing, the PCR products were amplified using primers CIITA-GM1F and CIITA-SEQR, 5′-ACAATCTCRAAACCTCRATTCTC-3′, and cloned into pCR4 vector using a TOPO-TA cloning Kit (Invitrogen), after which the plasmid DNA was purified using a PI system (Kurabo, Tokyo, Japan).

    Produced:

    Article Title: Characterization of HCV Interactions with Toll-Like Receptors and RIG-I in Liver Cells
    Article Snippet: The expression vector pTOPO was from Invitrogen (Carlsbad, CA) and the negative control siRNA was produced by T7 polymerase in vitro with the Ambion Silencer siRNA construction Kit or purchased (both control and kit from Ambion, Austin, TX). .. Core, Envelope E1/E2 and NS3/4A were amplified from pJFH-1 plasmid and cloned using TOPO TA Cloning kit from Invitrogen ( shows PCR primers used for amplification).

    Staining:

    Article Title: Calcium Influx and DREAM Protein Are Required For GnRH Gene Expression Pulse Activity
    Article Snippet: The PCR amplified products were separated by 1.0% TAE agarose gel electropheresis and visualized by ethidium bromide staining. .. The amplified RT-PCR fragment DAM12 (852 bp) was cloned into pCR2.1-TOPO vector using the TOPO TA Cloning Kit (Invitrogen).

    Article Title: Inactivation of class II transactivator by DNA methylation and histone deacetylation associated with absence of HLA-DR induction by interferon-γ in haematopoietic tumour cells
    Article Snippet: After ethanol precipitation, the DNA was subjected to 3% agarose gel electrophoresis and stained with ethidium bromide. .. For bisulphite sequencing, the PCR products were amplified using primers CIITA-GM1F and CIITA-SEQR, 5′-ACAATCTCRAAACCTCRATTCTC-3′, and cloned into pCR4 vector using a TOPO-TA cloning Kit (Invitrogen), after which the plasmid DNA was purified using a PI system (Kurabo, Tokyo, Japan).

    Fluorescence In Situ Hybridization:

    Article Title: In vivo dissection of the chromosome condensation machinery
    Article Snippet: .. PCR cloning was performed using the TOPO-TA cloning kit (Invitrogen). rDNA and CEN16 proximal FISH probes were generated as previously described ( ). .. Antibodies used were as follows: mAb YOL1/34 (rat anti-tubulin; Serotec), mAb 12CA5 (anti-HA; BAbCo), mouse antidigoxigenin and pig anti–goat-FITC (Boehringer), goat anti–mouse-FITC (BAbCo), rabbit anti-GFP (CLONTECH Laboratories, Inc.), and goat anti–rat-FITC and CY3 anti–rabbit (Jackson ImmunoResearch Laboratories).

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    Thermo Fisher topo xl pcr cloning kit
    Topo Xl Pcr Cloning Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 213 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher topo xl cloning kit
    Topo Xl Cloning Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/topo xl cloning kit/product/Thermo Fisher
    Average 90 stars, based on 25 article reviews
    Price from $9.99 to $1999.99
    topo xl cloning kit - by Bioz Stars, 2020-02
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