Qiagen
RNeasy MinElute Cleanup Kit (50)
74204
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I have used RNeasy MinElute Cleanup Kit for purification of rRNA depleted gram negative total RNA. Recovery was almost 80% with total input. After purification, the quantity was checked by qubit HS RNA assay and we got 13 to 16 ng/microliter.
Purification of total RNA for RNA-seq
rRNA depleted total RNA
80 microliters of rRNA depleted total RNA was made up to 100 microliter volume using 20 microliters of RNase free water. Then the entire 100 microliter mix was applied to Minelute spin columns and followed with lysis and washing using respective buffers provided in the kit. The protocol is very easy-to-follow and in less than 30 minutes, you will get high quality pure RNA which is ready for further down stream application like RNA-seq
As much as possible, avoid keeping your RNA at room temperature. In between steps, RNA should sit on ice.
Recovery was very good with good quality as checked in 2% agaorse gel. Got 13 to 15 ng per microliter after final elution in 12 microliter of RNase free water.
None
Very easy to perform protocol. It takes less than 30 minutes to perform purifciation.
Ideal kit for purification of total RNA in preparation of RNA-seq library pipeline.
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