RNA reverse transcription (RT) was performed according to Genomic DNA Eraser Reverse Transcription Kits (Takara, catalog no. RR047A). Briefly, 1 μg of RNA was treated with DNA eraser, and then cDNA was synthesized using PrimeScript RT enzyme mix and RT primer mix. To determine the GFP RNA transcript copy number, 100 ng RNA were subjected to real-time PCR.
Genomic dna eraser reverse transcription kits
Manufactured by Takara Bio
The Genomic DNA Eraser Reverse Transcription Kits are designed to remove genomic DNA and perform reverse transcription in a single step. The kits include reagents for efficient removal of genomic DNA contamination and subsequent cDNA synthesis from RNA samples.
Automatically generated - may contain errors
3 protocols using genomic dna eraser reverse transcription kits
1
cDNA Synthesis and qPCR Quantification
2
RNA Extraction and qPCR Quantification
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Total RNA was extracted with TRIzol reagent (Life Technologies) according to the manufacturer’s instructions. One microgram of RNA was used for reverse transcription with Genomic DNA Eraser reverse transcription kits (Takara). Quantitative real-time PCR was performed with SYBR select master mix (Life Technologies) according to the manufacturer’s instructions. Total DNA was extracted by phenol–chloroform extraction, and the copy number of viral genomes was measured as described previously (49 (link)).
3
Reverse Transcription and qPCR for GFP Quantification
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The mRNA was reverse-transcribed to cDNA according to Genomic DNA Eraser Reverse Transcription Kits (Takara, catalog no. RR047A). Briefly, 1 μg of RNA was treated with DNA eraser enzyme to remove genomic DNA, and then cDNA was synthesized using PrimeScript RT enzyme mix and RT primer mix. 100 ng RNA were subjected to Real-time PCR (Quantstudio 7, Applied Biosystems) to determine the GFP RNA transcript copy number, and the primers for RT-PCR were designed according to Gong et al.20 (link).
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