Cells-to-cDNA Kit is Not Easy as Promised

January 21, 2014

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review
King Abdullah Intl Med Res Ctr
Medical Genomics
Research Scientist

Company:

Ambion

Product Name:

Cells-to-cDNA II Kit

Catalog Number:

AM1722

Cells-to-cDNA kit is designed to minimise steps in carrying out conventional end point PCR. It has two options, One step and two step RT-PCR. In One step, you can only carry out endpoint PCR whereas in two step, the cDNA can be used for other applications. However, the overall process of optimization with Armored RNA control in pilot experiments did not work very well with me. Also, it remains to be seen if this kit can work with homogenized tissue from different sources.

Experimental Design and Results Summary

Application

Carrying out RT-PCR directly from Cells

Starting Material

Cultured Cells

Protocol Overview

The protocol steps are bit confusing with the use of Armored RNA control in pilot experiment. Regular process of lysis, chopping DNA, reverse transcription followed by PCR is experienced entangled in several incubation steps at different temperatures.

Tips

Use Two step RT-PCR unless you have real pressing issues of time. This allows more flexibility in using the cDNA

Results Summary

The use of conventional RNA extraction and cDNA preparation kits does not feel bad after using this kit which promises ease but actually does not.I worked with HCT-8 cells and used random primers for preparing cDNA. RT-PCR was done using CTNNB and GAPDH primers. It did not work. The Armored RNA control did work in the pilot study but the controls were not satisfactory.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

This kit does not have an expiry date.

The Bad

Did not fulfill the promise of easy steps and involves a lot of optimization based on arbitrary controls

The Bottom Line

This kit is useful if it is fully optimized and used in highthroughput applications. For a few cell lines and genes, I would still prefer separate kits for extraction and cDNA preparation

Join the discussion