The preparation and evaluation of AMs was conducted routinely. Total RNA was extracted using TRIzol and miRNeasy Mini Kit (Qiagen, Hilden, Germany) following the manufacturer’s protocol, except that isopropanol was replaced with ethanol for RNA precipitation. RNA quality was ascertained using 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). Then 1 μg of total RNA was reverse transcribed, and the product (11 μL) was preamplified using Megaplex PreAmp Primers and DBI Bestar® qPCR RT Kit (Applied Biosystems, Foster City, CA, USA) in a 20 μL PCR. The preamplification cycling conditions were 37°C for 60 minutes and 98°C for 10 minutes. The preamplified cDNA was diluted with 0.1× Tris-EDTA buffer (pH 8.0) to 10 μL, and then 1 μL of diluted cDNA was used in each plate for real-time PCRs (RT-PCRs).
Dbi bestar qpcr rt kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The DBI Bestar® qPCR RT Kit is a molecular biology tool designed for reverse transcription and real-time quantitative PCR (qPCR) analysis. It contains the necessary reagents and enzymes for both steps of the process.
Automatically generated - may contain errors
Lab products found in correlation
Megaplex preamp primers, by Thermo Fisher Scientific (2 mentions)
2100 bioanalyzer, by Agilent Technologies (1 mentions)
Mirneasy mini kit, by Qiagen (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions)
2 protocols using dbi bestar qpcr rt kit
1
Extraction and Evaluation of Amplified RNAs
2
Sensitive RNA Extraction and qRT-PCR
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Total RNA was harvested using Trizol (Invitrogen, CA, US) and an RNeasy Mini Kit (Qiagen, German) according to the manufacturer’s instructions. RNA quality was ascertained using an Agilent 2100 bioanalyzer (Agilent technologies). 1 μg of total RNA was reverse-transcribed and the product (11 μl) was pre-amplified using Megaplex PreAmp Primers and DBI Bestar® qPCR RT Kit (Applied Biosystems) in a 20 μl PCR reaction. The pre-amplification cycling conditions were 37 °C for 60 min and 98 °C for 10 min. The pre-amplified cDNA was diluted with 0.1 × TE (pH 8.0) to 10 μl and then 1 μl diluted cDNA was used in each plate for qRT PCR reactions.
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