Taq rt pcr kit

Manufactured by New England Biolabs

Sourced in Germany

The Taq RT-PCR Kit is a reagent system designed for reverse transcription and real-time PCR amplification of RNA targets. It contains Taq DNA polymerase, necessary buffers, and other reagents required for the RT-PCR process.

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Lab products found in correlation

Nanodrop 1000, by Avantor (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) Fast real time pcr system, by Thermo Fisher Scientific (1 mentions) Maxima sybr green rox qpcr master mix kit, by Thermo Fisher Scientific (1 mentions) Nanodrop 1000 spectrophotometer, by Thermo Fisher Scientific (1 mentions) Rnase h, by New England Biolabs (1 mentions)

Spelling variants of this product

Taq PCR Kit (14 citations) RT-PCR kit (3 citations)

2 protocols using taq rt pcr kit

RNA, DNA, and Protein Extraction from Explants

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Cells from explant cultures were lysed and processed with an RNA/DNA/Protein isolation kit (Isolate II, Bioline, London, UK) according to the manufacturer’s instructions. RNA quantity was determined using UV/VIS spectrophotometry (Nanodrop 1000, PeqLab, Erlangen, Germany). Protein concentration was analyzed with a Bradford Kit (Merck, Darmstadt, Germany). One Taq RT-PCR Kit (New England Biolabs INC, Frankfurt, Germany) was used to convert total RNA to cDNA with M-MulV Enzyme Mix and oligo dT primers. We used 500 ng of total RNA for one cDNA reaction.

Shi L., Stachon T., Käsmann-Kellner B., Seitz B., Szentmáry N, & Latta L. (2020). Keratin 12 mRNA expression could serve as an early corneal marker for limbal explant cultures. Cytotechnology, 72(2), 239-245.

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Quantifying Extracellular Vesicle mRNA Expression

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RNA was extracted using RNeasy mini kit (QIAGEN, Germany), and quantified by NanoDrop 1000 Spectrophotometer (Thermo Fisher, Wilmington, DE). cDNA was synthesized from RNA, using M-MuLV reverse transcriptase [Taq® RT-PCR kit, New England Biolabs (NEB), Ipswich, MA]. To remove any remaining RNA, the cDNA was treated with RNase H (NEB). qPCR was performed [Maxima SYBR Green/ROX qPCR Master Mix kit (Thermo Fisher) and 7500/7500 Fast Real-Time PCR System (Applied Biosystems, East Lyme, CT)], using GAPDH mRNA as endogenous control. The following primers were used: HChrR6 Forward 5′-GCAGATCCTCGTGTTCCTGGA-3′, HChrR6 Reverse 5′-CCTGGTCAATCACTTCTCCGTTCT-3′, GAPDH Forward 5′-GGGTGTGAACCATGAGAAGT-3′ and GAPDH Reverse 5′-GGCATGGACTGTGGTCATGA-3′. The EV mRNA copy number was estimated employing a standard curve, using the following formula: [X(ng)×6.0221×10²³ (molecules/mole)]/[N×330 (g/mole)×10⁹ (ng/g)], where X is the amount of EV mRNA, N is its length, and 330 is the average molecular weight of individual nucleotides (Rhode Island Genomics and Sequencing Center, Kingston, RI).

Wang J.H., Forterre A.V., Zhao J., Frimannsson D.O., Delcayre A., Antes T.J., Efron B., Jeffrey S.S., Pegram M.D, & Matin A.C. (2018). Anti-HER2 scFv-directed extracellular vesicle-mediated mRNA-based gene delivery inhibits growth of HER2-positive human breast tumor xenografts by prodrug activation. Molecular cancer therapeutics, 17(5), 1133-1142.

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