We used the RNAprep FastPure Tissue & Cell Kit (Tsingke Biotechnology, Beijing, China) and ABScript III Reverse Transcriptase (ABclonal, Wuhan, China) to extract messenger RNA (mRNA) from LUAD and normal tissue and performed reverse transcription according to the manufacturer’s protocol. Finally, qRT-PCR experiments were carried out using ABScript II One-Step SYBR Green RT-qPCR Kit (ABclonal). The primer sequences were as follows: CENPW: 5'-GAT GGA ACT GGC TGA GAC ACT AAC C-3' (forward) and 5'-AAG ACT CTT GCT TGA TGC TGA GGT G-3' (reverse); CENPM: 5'-ACA GCA AAT ACA GTC TCC AGA A-3' (forward) and 5'-GAA ACA CAC CTT CCC CAA GAA-3' (reverse); CENPU: 5'-GAA AAG AAA AGG CAG CGT ATG A-3' (forward) and 5'-AAT ATG CTG CAT TCC TAA GGG A-3' (reverse); CENPF: 5'-TAC AAC GAG AGA GTA AGA ACG C-3' (forward) and 5'-CTA CCT CCA CTG ACT TAC TGT C-3' (reverse); CENPH: 5'-TTC CAG AAC CTT ATT TTG GGG A-3' (forward) and 5'-CTT CTC AAG CTG CAG AAC AAT T-3' (reverse).
Rnaprep fastpure tissue cell kit
Manufactured by Tsingke
The RNAprep FastPure Tissue & Cell Kit is a nucleic acid extraction kit designed to purify total RNA from a variety of sample types, including tissues and cells. The kit utilizes a simple, spin-column-based protocol to efficiently isolate high-quality RNA that is suitable for downstream applications such as RT-PCR, real-time PCR, and RNA sequencing.
Automatically generated - may contain errors
2 protocols using rnaprep fastpure tissue cell kit
1
Quantification of CENP Genes in LUAD
2
Quantitative Reverse Transcription PCR Protocol
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To carry out quantitative reverse transcription PCR, total RNA was extracted from the cells using RNAprep FastPure Tissue&Cell Kit (Tsingke Biotechnology Co., Ltd). Reverse transcriptions were performed with Transcriptor cDNA Synthesis Kit (Roche, Basel, Switzerland) according to the manufacturer's instruction. The obtained cDNA was detected by Applied Biosystems™ 7500 with PerfectStart® Green qPCR SuperMix (TransGen, Beijing, China). The primers used in this study were as follows: ACE2 forward 5′- CAAGAGCAAACGGTTGAACAC-3′, reverse 5′- CCAGAGCCTCTCATTGTAGTCT-3’; SKP2 forward 5′-TTTCATGGGACTCCCTTCCG-3′, reverse 5′-GAGACAGTATGCCGTGGAGG-3’; GAPDH forward 5′-ACCACAGTCCATGCCATCAC-3′, reverse 5′-TCCACCACCCTGTTGCTGTA-3’. Target gene expression was normalized to that of GAPDH and determined by the comparative Ct method (ΔΔCT).
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