Our lab is using the Gene jet miniprep kit for plasmid Isolation. Somehow, the resuspension solution or alkaline lysis I buffer or Buffer p1 is going to empty soon even though we still have a lot of columns and other buffers left. I can make the resuspension buffer in the lab myself, but the question is what to do about the Rnase A to be added to the buffer.
How much volume and concentration of Rnase A is required for 100ml resuspension buffer ?
Hello
Look At the answer in these links
Good Luck
http://2014.igem.org/wiki/images/b/ba/Miniprep_protocol.pdf
http://www.genscript.com/site2/document/13326_20100901221715.PDF
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4972323/pdf/pone.0160509.pdf
I have prepared resuspension solution ( Tris 25mM, EDTA 10mM, Glucose 50mM) for plasmid isolation by using Gene Jet mini prep kit. I wish to add RNase A in this solution. I have Invitrogen purelink RNase A 20 mg/ml.
What concentration of RNaseA should I add in above mentioned resuspension solution?
Hi,
As a thumb rule 10-20 microliter of RNase A (20 mg/ml) may be used for the bacterial sample having OD-1, by that you can calculate, how much volume of lysis buffer you are using for the resuspension. In general, 20 microliter RNase A can be added to 200 microliter lysis buffer.
Good Luck.
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Nilesh
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