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Old 06-05-2012, 01:33 PM   #1
Eli Boritz
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Default oligo-dT Dynabeads wash buffer for mRNA-Seq

The Dynabeads protocol calls for a wash buffer that includes Lithium chloride, but other published protocols (ex., http://www.ncbi.nlm.nih.gov/pubmed/21807852) replace LiCl with NaCl. I presume this is because Lithium might inhibit RT, but am not sure. Does anyone who does a lot of Illumina TruSeq-based mRNA-seq have a preferred wash buffer for the oligo-dT beads?
Thanks very much!
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Old 06-11-2012, 07:12 AM   #2
mikaelk
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In the lab we are using a directional mrna-seq protocol based on illumina tru-seq smallrna kits. I have not compared the two types of buffers but I have been doing polyA purifications with LiCl based buffers and I have still been able to construct libraries without any problem. Hope this helps a little.
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Old 06-11-2012, 08:08 AM   #3
protist
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We have used the Li containing buffers and have had no issues with libraries - even from small amounts of starting material.
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