Taqman reverse transcription reagent kit

Manufactured by Takara Bio

Sourced in Japan

The Taqman Reverse Transcription reagent kit is a set of reagents used for the reverse transcription of RNA to cDNA. The kit includes essential components necessary for the reverse transcription reaction, such as reverse transcriptase enzyme, buffer, and primers.

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Lab products found in correlation

Sybr green pcr master mix, by Takara Bio (1 mentions) Real time pcr system, by Takara Bio (1 mentions) Rneasy kit, by Qiagen (1 mentions) Anti mettl3, by Abcam (1 mentions) Anti caprin 1, by Proteintech (1 mentions) Anti wtap, by Santa Cruz Biotechnology (1 mentions) Anti hif 1α, by Proteintech (1 mentions) Sybr green master mix, by Takara Bio (1 mentions) Anti gapdh, by Cell Signaling Technology (1 mentions)

2 protocols using taqman reverse transcription reagent kit

Liver RNA Extraction and qPCR Analysis

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Liver was homogenized and total RNA was isolated using the RNeasy kit (Qiagen, Hilden, Germany). A reverse transcription reaction was performed by Taqman Reverse Transcription reagent kit (Takara, Kusatsu, Japan) according to the manufacturer's instructions. The real-time PCR reaction was carried out on Roche real-time PCR system with cDNA sample containing SYBR Green PCR master mix (Takara) and primers. Primer sequences can be made available upon request. Relative expression of mRNA was calculated by the comparative cycle threshold method.

Xing Y.W., Lei G.T., Wu Q.H., Jiang Y, & Huang M.X. (2019). Procyanidin B2 protects against diet-induced obesity and non-alcoholic fatty liver disease via the modulation of the gut microbiota in rabbits. World Journal of Gastroenterology, 25(8), 955-966.

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Quantitative RT-PCR and Western Blot Analysis of mRNA and Protein Expression

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For mRNA analysis, total RNA extraction for cDNA synthesis was conducted with the Taqman Reverse Transcription Reagent kit (TaKara-Bio, Kusatsu, Japan). Real-time quantitative PCR was conducted with different primer sequences (Table 1) using SYBR Green Master Mix (TaKaRa).Table 1

Primer sequences for real time RT-PCR

Target	Forward	Reverse
ACTB	TCTTCCAGCCTTCCTTCCT	AGCACTGTGTTGGCGTACAG
CAPRIN1	TCTCGGGGTGATCGACAAGAA	CCCTTTGTTCATTCGTTCCTGG
SLC2A1	TGTCTGGCATCAACGCTGTCTTC	TC CCTGCTCGCTCCACCACAAAC
HK2	CGACAGCATCATTGTTAAGGAG	CA GCAGGAAAGACACATCACATTT
HIF1A	AGTTCCGCAAGCCCTGAAAGC	GCAGTGGTAGTGGTGGCATTAGC
MYC	CGCCTCTTGACATTCTCCTC	GGACTATCCTGCTGCCAAGA
NUP160	GTTATCTGGCTGCTCTCAATTG	GTGCATTCTCCATCATGATTCC
NUP133	AGTACCTGTGGGCTGCTTCTCTAG	GCTCTGGTTGTCAGTCTGCTCAC
NUP155	CCGCTCCTCAGTCTCCCAGTG	GCTCATCCTTGGATCGCTGTGAC
METTL3	CCAGCACAGCTTCAGCAGTTCC	GCGTGGAGATGGCAAGACAGATG
WTAP	CTGACAAACGGACCAAGTAATG	AAAGTCATCTTCGGTTGTGTTG

Proteins were separated by SDS–PAGE followed by Western blot as described previously [30 (link), 31 (link)]. Anti- GAPDH (Cell Signaling Technology), anti-Caprin-1 (116 kDa, ProteinTech Group), anti-HIF1α (120 kDa, ProteinTech Group), anti-c-Myc (49 kDa, roteinTech Group), anti-WTAP (45 kDa, Santa Cruz Biotechnology), and anti-METTL3 (64 kDa, Abcam) were used as primary antibodies. HRP-conjugated anti-mouse and anti-rabbit IgG (Cell Signaling Technology) were used as secondary antibodies.

Gao Y., Yuan L., Ke C., Pei Z., Liu X., Wu R., Kui X, & Zhang Y. (2023). Caprin-1 plays a role in cell proliferation and Warburg metabolism of esophageal carcinoma by regulating METTL3 and WTAP. Journal of Translational Medicine, 21, 159.

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