Real-time qPCR was performed using the SYBR qPCR detection Kit (TaKaRA, Tokyo, Japan) and the Real-time PCR System (Roche, Basel, Switzerland). PCR was performed in the relative quantification of mRNA expression using the comparative cycle threshold (CT) method. 18S expression was used as the endogenous control. The primers used were listed in Supplementary Table 1 .
Sybr qpcr detection kit
Manufactured by Takara Bio
Sourced in Japan, China
The SYBR qPCR Detection Kit is a reagent designed for real-time quantitative PCR (qPCR) analysis. It contains SYBR Green I, a fluorescent dye that binds to double-stranded DNA, enabling the detection and quantification of target DNA sequences during the PCR amplification process.
Automatically generated - may contain errors
2 protocols using sybr qpcr detection kit
1
Quantitative Real-time PCR for mRNA
2
Quantitative RT-PCR Analysis of Gene Expression
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Total RNA was extracted from cells using Trizol reagent (Invitrogen). RNA was reversely transcribed into cDNA using PrimeScript RT Master Mix (TaKaRa, Dalian, China). The cDNA was subjected on Real-time polymerase chain reaction (PCR) analysis using the SYBR qPCR-detection-Kit (TaKaRA, Dalian, China) on the Real-time PCR System (ABI7500, ABI, Oyster Bay, NY, USA). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the internal control. The sequences of the primers used for RT-PCR were listed in Table 2 .
Primers sequences for qRT-PCR
| Genes | Primer sequences |
|---|---|
| CARM1 | F: 5′-TTGATGTTGGCTGTGGCTCTGG-3′ |
| R: 5′-ATGGGCTCCGAGATGATGATGTCC-3′ | |
| MMP24 | F:5′-CTCTGGCCAGTGCTCACC-3′ |
| R:5′- CCCATAATTCCCTGCCCCAC-3′ | |
| MAP7 | F:5′-CAGTGCGAAGCGAAACAGC-3′ |
| R:5′-TGTTTCTCCCGTTCCTCACG-3′ | |
| ZFP91 | F:5′-GCATGGGACAGCTCAGACTT-3′ |
| R:5′-GTTGGACGATCGGGTTGGAA-3′ | |
| ANKH | F:5′-CATCGGAGTGGACTTTGCCT-3′ |
| R:5′-GATCAGGAAGGGCATACCCAG-3′ | |
| LASP1 | F:5′-CCCAAGCAGTCCTTCACCAT-3′ |
| R:5′-AGGAATCACAAGCTGTCGCA-3′ | |
| ATG2A | F:5′-TGCCAATCTGCTGTGAGAGG-3′ |
| R:5′-ACGCACTCACGGAGCTTAAA-3′ | |
| HSPA8 | F:5′-CCCCATCATCACCAAGCTGT-3′ |
| R:5′-CTCCACCACCAGGAAATCCC-3′ | |
| ULK1 | F:5′-CCTGAGGAGACCCTCATGGA-3′ |
| R:5′-CACAGCTTGCACTTGGTGAC-3′ | |
| CCNG2 | F:5′-AACAAAAACAAGGGGCTCGG-3′ |
| R:5′-ATCATTCTCCGGGGTAGCCT-3′ | |
| GAPDH | F: 5′-CTCTGATTTGGTCGTATTGGG-3′ |
| R: 5′-TGGAAGATGGTGATGGGATT-3′ |
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