Total RNA was extracted from tissues or cultured cells through Trizol using the TransZol Kit (TransGen Biotech, Beijing, China) and reverse transcribed using the NovoScript®Plus All-in-one 1st Strand cDNA Synthesis SuperMix (Novoprotein, Shanghai, China). The concentration and purity of the cDNA were determined using Varioskan LUX (Thermo Fisher, USA), and real-time quantitative PCR was performed on a Bio-Rad CFX96 Touch Real-Time PCR Derection System (Bio-Rad, Hercules, CA, USA) using the NovoStart® Fast SYBR qPCR SuperMix (Novoprotein, China). Primers were obtained from Sangong Biotechnology Co., Shanghai, China, and the sequences are shown in Table 2 .
Novostart fast sybr qpcr supermix
Manufactured by Novoprotein
Sourced in United States
NovoStart® Fast SYBR qPCR SuperMix is a ready-to-use solution for real-time quantitative PCR (qPCR) reactions. It contains all the necessary components, including a fast-acting DNA polymerase, SYBR Green I dye, and optimized buffer, to enable rapid and efficient amplification and detection of target DNA sequences.
Automatically generated - may contain errors
Lab products found in correlation
Novoscript plus all in one 1st strand cdna synthesis supermix, by Novoprotein (2 mentions)
Nanodrop, by Thermo Fisher Scientific (2 mentions)
Transzol kit, by Transgene (1 mentions)
Varioskan lux, by Thermo Fisher Scientific (1 mentions)
Abi steponeplus instrument, by Thermo Fisher Scientific (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
2 protocols using novostart fast sybr qpcr supermix
1
RNA Extraction and qPCR Analysis Protocol
2
Quantitative RT-PCR of Liver RNA
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Total RNA from liver was isolated using TRIzol (Invitrogen, Carlsbad, CA) method. The concentration and purity of total RNA were determined using Nanodrop (ND 2000, Thermo, Waltham, MA). Sample selection was based on the criteria that the OD260/OD280 ratio of total RNA should fall within the range of 1.8 to 2.0, and the OD260/OD230 ratio should be between 2.0 and 2.2. The cDNA synthesized by using NovoScript Plus All-in-one 1st Strand cDNA Synthesis SuperMix (Novoprotein:E047-01B), the Reverse Transcription Reaction System was showed in Supplementary Table 1 . qRT-PCR was carried out using NovoStart Fast SYBR qPCR SuperMix (Novoprotein:E301-01A), the specific steps are as follows: the first step is predenaturation, 95℃, 30 s, the number of cycles is 1; The second step is denaturation and annealing/extension stage, denaturation 5 s at 95℃, annealing/extension 30 s at 60℃, PCR reaction cycle 40 times, reaction system is showed in Supplementary Table 2 . Primers were designed by Primer 5, synthesized by Servicebio (Wuhan, China) and the Primer information was showed in Supplementary Table 3 . Results were analyzed on an ABI Step One Plus instrument (Applied Biosystems, Carlsbad, CA) using the method of 2−△△Ct.
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