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Old 12-04-2012, 11:23 PM   #1
megalodon
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Location: Pennsylvania

Join Date: Sep 2010
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Default Amplification of Bacterial total RNA

Hi,

I am looking for an amplification protocol for bacterial RNA that works with very small amounts of RNA (low ng- to pg-range). I need to amplify RNA that was extracted from very small environmental samples to get enough starting material for the preparation of Illumina sequencing libraries.

Has anyone amplified bacterial RNA prior to next-gen library preparation? My understanding is that amplifying bacterial RNA is not straight forward due to the lack of a poly(A) tail. The few kits that I found (e.g. Ambion MessageAmp II - Bacterial Kit) are mostly used for microarray-based whole genome expression profiling and these kits still need a min. of 10ng of RNA for amplification.

What is the best way of turning a very small amount of bacterial RNA into an Illumina library?

Any help with this would be greatly appreciated!

Thanks a bunch in advance!

Daniela
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