All the reactions were performed in a 20-μL reaction volume in triplicate by SYBR Green Real-time PCR Universal Reagent (GenePharma Co., Ltd.) and MX-3000P Real-time PCR equipment (Stratagen). Standard curves were generated, and the relative amount of ALDH1 was normalized to 18S RNA (2−ΔΔCt). The ALDH1 expression fold change was evaluated using 2−ΔΔCt. The primers are ALDH1 (forward, 5′-CTGCTGGCGACAATGGAGT-3′; reverse, 5′-GTCAGCCCAACCTGCACAG-3′) and 18S rRNA (forward 5′-CCTGGATACCGCAGCTAGGA-3′ and reverse 5′-GCGGCGCAATACGAATGCCCC -3′). These primers yielded 230- and 112-bp products, respectively.
Sybr green real time pcr universal reagent
Manufactured by GenePharma
SYBR Green Real-time PCR Universal Reagent is a laboratory product designed for use in real-time polymerase chain reaction (qPCR) experiments. It contains SYBR Green I dye, which binds to double-stranded DNA and emits fluorescence, allowing for the quantification of DNA amplification during the qPCR process.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using sybr green real time pcr universal reagent
1
ALDH1 Expression Quantification by qPCR
2
Quantification of miRNA-100-5p Expression
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The RNAs were collected from cells by Trizol extraction (Invitrogen), and cDNAs were acquired using an RNA reverse transcription amplification kit (Takara). RT-PCR assays were performed by SYBR Green Real-time PCR Universal Reagent (GenePharma Co.Ltd.) and analyzed by BIO-RAD fluorescence quantitative PCR machine. Primers were as follows:
miR-100-5p (5′-AACCCGTAGATCCGAACTTGTG-3′);
u6 (5′-ACGCAAATTCGTGAAGCGTT-3′);
GAPDH (forward 5′-TGGAAATGACAGTGAAGCACCTC-3′.
reverse 5′-TCGTTCATGCACTCGCTGAAG-3′);
3
Quantitative Analysis of miRNA-100-5p
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The RNAs were collected from cells by Trizol extraction (Invitrogen), and cDNAs were acquired using an RNA reverse transcription ampli cation kit (Takara). RT-PCR assays were performed by SYBR Green Realtime PCR Universal Reagent (GenePharma Co.,Ltd.) and analyzed by BIO-RAD uorescence quantitative PCR machine. Primers were as follows: miR-100-5p (5'-AACCCGTAGATCCGAACTTGTG-3'); u6(5'-ACGCAAATTCGTGAAGCGTT-3'); GAPDH(forward5'-TGGAAATGACAGTGAAGCACCTC-3' reverse5'-TCGTTCATGCACTCGCTGAAG-3'); u6 was used as control for miRNA, GAPDH was used as control for mRNA. Both of them was calculated with the 2 - ΔΔCT method.
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