cDNA synthesis by RevertAid First Strand cDNA Synthesis Kit

University of Tennessee Health Science Centre
Pathology
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

Thermo Scientific

Product Name:

RevertAid First Strand cDNA Synthesis Kit

Catalog Number:

K1622

Thermo Scientific RevertAid First Strand cDNA Synthesis Kit is a complete system for efficient synthesis of first strand cDNA from RNA templates. Total RNA was isolated from MCF7 cells. 1 microgram of total RNA was used as input for cDNA synthesis. The cDNA was used as template for desired gene amplification with specific primers by PCR.

Experimental Design and Results Summary

Application

Reverse Trancription - cDNA synthesis

Starting Material

Total RNA

Protocol Overview

one microgram of isolated total RNA was taken as input. All components of RT reactions such as Random Hexamer, 5 X Reaction buffer, dNTP, RNAse inhibitor, Reverse transcriptase enzyme (M-MuLV RT) weres added according to kit instruction. The total reaction volume was 20 microlitre and the reaction mixture was incubated for 5 min at 25°C followed by 60 min at 42°C.Reaction was terminated by heating at 70°C for 5 min. The reverse transcription reaction product can be directly used in PCR applications or stored at -20°C for future use.

Tips

RNase free environment is essential - clean the working area with DEPC

Results Summary

The kit produced good quality cDNA, which was confirmed by good PCR amplification of desired gene.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Efficient cDNA synthesis kit

The Bad

Need to use high amount of input

The Bottom Line

The kit produced good quality of cDNA, which was confirmed by gene amplification of desired gene by PCR.

Join the discussion