Methanol and acetonitrile were of MS grade from Merck (Darmstadt, Germany). Formic acid (MS grade), hexafluoro-2-propanol (HFIP, ≥99.0%), trimethylamine (TEA, ≥99.5%), Transcriptor Universal cDNA Master kit, protease Inhibitor Cocktail, phosphatase inhibitor, phosphodiesterase I and II from Crotalus adamanteus venom were purchased from Sigma-Aldrich (St. Louis, MO, USA). TRIzol reagent, mirVana™ miRNA isolation kit, Pierce™ Streptavidin-coated magnetic beads, RNase I and RNase T1 were purchased from Thermo Fisher Scientific (Waltham, MA, USA). A low-range ssRNA ladder and shrimp alkaline phosphatase were purchased from New England Biolabs (Beverly, MA, USA). RPMI-1640 medium, fetal bovine serum (FBS), penicillin, streptomycin and puromycin were obtained by Gibco (Carlsbad, CA, USA). RNase A was purchased from Takara Biotechnology (Dalian, China). Reference standards, 4-demethylwyosine (imG-14), wyosine (imG) and isowyosine (imG2), were provided by 9dingchem Co., Ltd (Shanghai, China). The purity was more than 98%. Taxol (purity ≥ 98%) was supplied by AdooQ bioscience (Nanjing, China). Ultrapure water (18.25 MΩ·cm) was produced using a Milli-Q system from Millipore (Millipore, Bedford, MA, USA).
Transcriptor universal cdna master kit
Manufactured by Merck Group
Sourced in United States
The Transcriptor Universal cDNA Master kit is a laboratory equipment product designed for the reverse transcription of RNA to complementary DNA (cDNA). The kit provides the necessary reagents and components to perform this essential step in various molecular biology and gene expression analysis applications.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Rnase 1, by Thermo Fisher Scientific (1 mentions)
Rnase t1, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Milli q system, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Puromycin, by Thermo Fisher Scientific (1 mentions)
Trimethylamine, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Formic acid, by Merck Group (1 mentions)
Shrimp alkaline phosphatase, by New England Biolabs (1 mentions)
Pierce streptavidin coated magnetic beads, by Thermo Fisher Scientific (1 mentions)
Taxol, by Adooq (1 mentions)
Rnase a, by Takara Bio (1 mentions)
Hexafluoro 2 propanol, by Merck Group (1 mentions)
Nanodrop spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Mirneasy kit, by Qiagen (1 mentions)
Lightcycler 96, by Roche (1 mentions)
Faststart sybr green master kit, by Merck Group (1 mentions)
2 protocols using transcriptor universal cdna master kit
1
Optimized Extraction and Quantification of Modified RNA Nucleosides
2
Quantifying Gene Expression via RT-qPCR
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Cells were treated with SAF-Hex and cultured to extract mRNA using the miRNeasy kit (Qiagen, Germany). The mRNA quantity was measured using a NanoDrop spectrophotometer (Thermo Fisher). Briefly, 1 µg of total mRNA was reverse transcribed with the Transcriptor Universal cDNA Master kit (Sigma Aldrich, US). Real-time PCR reactions were performed using the FastStart SYBR Green Master kit (Sigma Aldrich), primers, and a 50 ng cDNA template. Primer sequences were a combination of previously established [13 (link), 14 (link)] and newly designed sequences (Table S2 ). qPCR was performed using a Light Cycler 96 instrument (Roche Diagnostics, Germany). The thermal cycling protocol comprised an initial denaturation step at 95 °C for 10 min, followed by 45 cycles of denaturation at 95 °C for 15 s, annealing at specified temperatures for 15 s, and extension at 72 °C for 20 s. Gene expression levels were normalized using the 2−ΔΔCt method, with GAPDH as the reference gene.
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