Regulatory regions (R1 and R2), identified by integration of previously published human islet ATAC-seq and H3K27ac ChIP-seq datasets [5 (link)], were PCR-amplified from genomic DNA extracted from EndoC-βH3 cells with primer sets (Table S5 ) designed by Primer3-based software and cloned into pGL4.23[luc/minP] vector (Promega) between KpnI and XhoI restriction enzyme sites. Plasmid DNA was extracted using mini-prep plasmid extraction kit and/or Maxi-prep plasmid extraction kit (QIAGEN). The constructs were further confirmed by the Sanger sequencing.
Pgl4.23 luc minp vector
Manufactured by Promega
The PGL4.23[luc/minP] vector is a plasmid designed for the expression of the luciferase gene under the control of a minimal promoter. It is a tool for studying gene regulation and transcriptional activity in various biological systems.
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2 protocols using pgl4.23 luc minp vector
1
Regulatory regions cloning and validation
2
Regulatory Region Cloning and Luciferase Assay
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Regulatory regions (R1 and R2), identified by integration of previously published human islet ATAC-seq and H3K27ac ChIP-seq datasets [11 (link)], were PCR-amplified from genomic DNA extracted from EndoC-βH3 cells with primer sets (Table S5 ) designed by Primer3-based software and cloned into pGL4.23[luc/minP] vector (Promega) between KpnI and XhoI restriction enzyme sites. Plasmid DNA was extracted using mini-prep plasmid extraction kit and/or Maxi-prep plasmid extraction kit (QIAGEN). The constructs were further confirmed by the Sanger sequencing.
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