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4 topo vector

Manufactured by Thermo Fisher Scientific
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The 4-TOPO vector is a DNA cloning vector used for the efficient insertion and expression of target genes in a variety of host organisms. It features a linearized backbone with single 3' thymine (T) overhangs, which facilitates direct ligation of PCR products amplified with compatible 3' adenine (A) overhangs. The vector also includes essential elements for selection and maintenance in bacterial hosts.

Automatically generated - may contain errors

Lab products found in correlation

Pyromark q24 platform, by Qiagen (2 mentions) Epitek bisulfite kit, by Qiagen (2 mentions)

Close spelling variants of this product

TOPO vector (87 citations) PcDNA3.4-TOPO Vector (6 citations) PCR-TOPO 4.0 vectors (3 citations) PCR®4-TOPO® vector system (2 citations)

2 protocols using 4 topo vector

1

Methylation Analysis of K+ Channel Genes

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To investigate the methylation pattern of CpG sites near the transcriptional start site (TSS) of four K+ channel genes, methylation data obtained from the L5 and L6 DRGs of control and SNL rats 3 weeks after surgery were compared. Genomic DNA obtained from pools of DRGs was treated with bisulfite using an Epitek bisulfite kit (Qiagen). This process converts the unmethylated cytosine to uracil but leaves the methylated cytosine unchanged. Using specific primers, the regions of interest close to the TSS of Kcna4, Kcnd2, Kcnq2, and Kcnma1 were amplified by PCR. The amplified DNA was cloned into 4-TOPO vector (Invitrogen), and the DNA obtained from individual clones after bacteria transformation was sequenced (Beckman Coulter Genomics). Each sequence, corresponding to a different allele, was analyzed for the presence of methylated cytosines using the QUMA web-based program (http://quma.cdb.riken.jp/). Primer sequences for bisulfate sequence are listed in Supplementary Table 4. Bisulfite-treated DNA obtained from the same samples was also analyzed by pyrosequencing technology using the PyroMarkQ24 platform (Qiagen).
Laumet G., Garriga J., Chen S.R., Zhang Y., Li D.P., Smith T.M., Dong Y., Jelinek J., Cesaroni M., Issa J.P, & Pan H.L. (2015). G9a Is Essential for Epigenetic Silencing of K+ Channel Genes in Acute-to-Chronic Pain Transition. Nature neuroscience, 18(12), 1746-1755.
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2

Methylation Analysis of K+ Channel Genes

Check if the same lab product or an alternative is used in the 5 most similar protocols
To investigate the methylation pattern of CpG sites near the transcriptional start site (TSS) of four K+ channel genes, methylation data obtained from the L5 and L6 DRGs of control and SNL rats 3 weeks after surgery were compared. Genomic DNA obtained from pools of DRGs was treated with bisulfite using an Epitek bisulfite kit (Qiagen). This process converts the unmethylated cytosine to uracil but leaves the methylated cytosine unchanged. Using specific primers, the regions of interest close to the TSS of Kcna4, Kcnd2, Kcnq2, and Kcnma1 were amplified by PCR. The amplified DNA was cloned into 4-TOPO vector (Invitrogen), and the DNA obtained from individual clones after bacteria transformation was sequenced (Beckman Coulter Genomics). Each sequence, corresponding to a different allele, was analyzed for the presence of methylated cytosines using the QUMA web-based program (http://quma.cdb.riken.jp/). Primer sequences for bisulfate sequence are listed in Supplementary Table 4. Bisulfite-treated DNA obtained from the same samples was also analyzed by pyrosequencing technology using the PyroMarkQ24 platform (Qiagen).
Laumet G., Garriga J., Chen S.R., Zhang Y., Li D.P., Smith T.M., Dong Y., Jelinek J., Cesaroni M., Issa J.P, & Pan H.L. (2015). G9a Is Essential for Epigenetic Silencing of K+ Channel Genes in Acute-to-Chronic Pain Transition. Nature neuroscience, 18(12), 1746-1755.
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