My group is planning to collect samples by buccal swab and extract DNA, RNA and protein. I have a few questions about collecting sample and method of extraction.
Can anyone share their experience on sampling and extraction? Thanks.
yes, you can extract RNA from buccal swab by RNasy kit and then storage it at -20.
Thank you Fathi. Maybe my question is not clear enough. Since it is a long term sample collection, so we want to accmulate certain number of buccal swabs before RNA extraction. After collecting the cells, can I directly store the swab at -80C or storing the swab in RNAlater is much prefered?
Yes you can. I would suggest to store your samples in -80 and when your are ready for extraction you can use the Qiagen kit for RNA and protein extraction.
After sample collection, place your swabs on ice for transportation and then put them immediately into a -80 freezer. Trizol extraction is better for the recovery of high purity RNA. You could use trizol following the liquid liquid extraction procedure or there ias a commerical kit by Ribopure that may be easier but I am not sure of the quality.
The most critical part is getting the sample into a stabilizing solution as early as possible to prevent RNA degradation and bacterial growth. This company has kits specifically for DNA, RNA, and Protein designed so that you use the swab and then scrape cells directly into a stabilizing solution so that you can store and ship samples at room temperature:
http://www.mawidna.com/products/iswab-rna-collection-kit/
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