Genomic DNA of HaCaT and HaCaT-T cells was extracted using Animal Genomic DNA Kit (TsingKe Biotech, China). Exons 5–9 of the p53 gene were amplified by PCR. The sequences of the primers are listed in Supplementary Table 2 . PCR was performed under the conditions: 94 °C for 3 min, 1 cycle, denaturation at 94 °C for 30 s, annealing at 58 °C for 3 s, extension at 72 °C for 1 min, 35 cycles. The PCR products of p53 gene in HaCaT and HaCaT-T cells were separated by gel electrophoresis in a 3% low melting agarose gel with EtBr. Subsequently, the PCR products were extracted from the gel, which was melted at 50 °C. The PCR products were reamplified under the same PCR conditions. The PCR products were then cloned into pTZ57R/T vector using InsTAclone PCR Cloning Kit (Thermo Scientific, Canada) and subject to DNA sequencing at the TsingKe Biotech Inc (Beijing, China).
Animal genomic dna kit
Manufactured by Tsingke
Sourced in China
The Animal Genomic DNA Kit is a laboratory tool designed for the extraction and purification of high-quality genomic DNA from a variety of animal samples. The kit utilizes a silica-based membrane technology to efficiently capture and isolate DNA, providing a reliable method for downstream genetic analysis and applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using animal genomic dna kit
1
Sequencing of p53 Gene Exons in HaCaT Cells
2
Quantifying Relative Telomere Length
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The relative human telomere length was quantified by measuring the amount of telomere repeat amplification product to a single copy gene product (IFNB1) using qPCR as described65 (link),70 (link). The genome DNA (gDNA) was isolated using the animal genomic DNA kit (Tsingke, Beijing) and quantified by spectrophotometer. 0.5–10 ng gDNA was used in qPCR with SYBR® Green Supermix (Bio-Rad, Catalog #1725121). The primers used were:
TEL-F, 5′-CGGTTTGTTTGGGTTTGGGTTTGGGTTTGGGTTTGGGTT-3′;
TEL-R, 5′-GGCTTGCCTTACCCTTACCCTTACCCTTACCCTTACCCT-3′;
IFNB1-F, 5′-GGTTACCTCCGAAACTGAAGA-3′;
IFNB1-R, 5′-CCTTTCATATGCAGTACATTAGCC-3′.
TEL-F, 5′-CGGTTTGTTTGGGTTTGGGTTTGGGTTTGGGTTTGGGTT-3′;
TEL-R, 5′-GGCTTGCCTTACCCTTACCCTTACCCTTACCCTTACCCT-3′;
IFNB1-F, 5′-GGTTACCTCCGAAACTGAAGA-3′;
IFNB1-R, 5′-CCTTTCATATGCAGTACATTAGCC-3′.
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