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Old 07-01-2010, 11:37 AM   #1
Paulo.jt
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Location: Brazil

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Default RNA cleanup after fragmentation - illumina

Does anybody know any alternative to RNA precipitation after the fragmentation step?
Ive been thinking about RNeasy MinElute Cleanup from Qiagen or SPRI beads. In this case, which beads are more adequate? Would AMPure XP work?

Thanks
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Old 07-01-2010, 12:09 PM   #2
tplsmith
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Default RNA ppt vs SPRI or MinElute

Either SPRI or MinElute frequently lower your yield; if RNA is not limiting this is not a problem. Recommend that you have calibrated your SPRI beads if they are XP as there is slight risk of selectively losing fragments in your target size range. Haven't actually used SPRI for this but have successfully used MinElute albeit common to only get 1/2 of the RNA back.
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Old 07-02-2010, 04:00 AM   #3
squeak
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Default

Just be aware that Ampure XP gets rid of all fragments of 100bp or less. I'm not sure what it is for the other methods.
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