Exon-Level Microarrays from Small Quantities of Tissue

June 17, 2019

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Company:

Affymetrix

Product Name:

WT pico Kit

Catalog Number:

902661

The whole transcriptome pico kit allows for small quantities of RNA from tissues to be processed to obtain a high yield of cDNA necessary for hybridization on Affymetrix human HTA arrays.

Experimental Design and Results Summary

Application

Microarrays

Starting Material

20-5000 pg total RNA

Protocol Overview

The protocol is completed in two days: on the first, cDNA is syntesized, then PCR amplified (6-12 cycles) to obtain enough templeate for the overnight in vitro translation. In the following day cRNA is purified, reverse-transcribed with modified nucleotides and purified from hydrolized cRNA. The final steps fragment and label the cDNA for use on Affymetrix microarrays.

Tips

When starting from very low amount of RNA quantification can be performed by qPCR with an appropriate housekeeping standard.

Results Summary

12 samples were processed. Of them, 11 yielded a usable microarray profile, while 1 gave very low signal, despite quantification of the probe was in the same range as the other samples.

Additional Notes

None

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Summary

The Good

Allows for extended gene profiling from minuscule amounts of total RNA.

The Bad

Involves amplification of the material, this could lead to distorted representation of transcripts.

The Bottom Line

Whole transcriptome profiling can be achieved by individual researchers through microarray platforms with no need for dedicated skills such as the deconvolution of RNAseq. The WT pico kit gives access to such technology even with hundreds of cells as starting material.

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