RNA extraction was performed using the RNA-Quick Purification Kit (ES Science, shanghai, China) following the manufacturer’s instructions. Reverse transcription was conducted according to Eco M-MLV RT Premix kit (AG11706, Accurate Biology, Atlanta, Georgia). Supplementary Table S1 shows the primer sequences used for RT-qPCR. Candidate gene expression was normalized to that of glyceraldehyde 3-phosphate dehydrogenase (GAPDH). RT-qPCR was performed using the SYBR Green Premix Pro Tag HS qPCR kit (AG11701, Accurate Biology) by QuantStudio 1 (applied biosystems, Thermo Fisher Scientific, Waltham, MA, USA). Samples were run and analyzed in triplicate.
Eco m mlv rt premix kit
Manufactured by Accurate Biology
The Eco M-MLV RT Premix kit is a reagent designed for the reverse transcription (RT) of RNA into complementary DNA (cDNA). It contains the necessary components, including the Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) enzyme, required for the RT reaction.
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Lab products found in correlation
3 protocols using eco m mlv rt premix kit
1
RNA Extraction and RT-qPCR Protocol
2
Profiling lncRNA Expression via RT-qPCR
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RNA extraction was performed using the RNA-Quick Purification Kit (ES Science) following the manufacturer’s instructions. Reverse transcription was conducted according to the Eco M-MLV RT Premix kit (AG11706, Accurate Biology). Target gene expression was normalized to the endogenous control gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH). RT-qPCR was conducted on the QuantStudio 1 (applied biosystems, Thermo Fisher Scientific, USA) machine utilizing SYBR Green Premix Pro Tag HS qPCR kit (AG11701, Accurate Biology). Expression levels of lncRNAs were calculated with 2−ΔΔCT method. Table 2
3
Quantitative RT-PCR Analysis of Gene Expression
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Total RNA was extracted using AG RNAex Pro Reagent (Accurate Biology, Cat#AG21102) following the manufacturer’s protocol. The RNA was reverse-transcribed using Eco M-MLV RT Premix kit (Accurate Biology, Cat#AG11706) and the quantification of RNA was detected using SYBR Green Pro Taq HS kit (Accurate Biology, Cat#AG11701). We performed the reactions on the Roche LightCycler 480II PCR instrument (Basel, Switzerland). The primer sequences are as follows:
GAPDH: forward primer (5’-GGAGCGAGATCCCTCCAAAAT-3’) and reverse primer (5’-GGCTGTTGTCATACTTCTCATGG-3’);
UCK2: forward primer (5’-AGATCCCCGTGTATGACTTTGT-3’) and reverse primer (5’-GGCTTGACGAACGTAATGTACTG-3’).
GAPDH: forward primer (5’-GGAGCGAGATCCCTCCAAAAT-3’) and reverse primer (5’-GGCTGTTGTCATACTTCTCATGG-3’);
UCK2: forward primer (5’-AGATCCCCGTGTATGACTTTGT-3’) and reverse primer (5’-GGCTTGACGAACGTAATGTACTG-3’).
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