Hi, is it possible to use the QIAquick PCR purification kit to purify RNA/cDNA hybrids from low-molecular fragments after RT without loosing the RNA strand? I need to work with the RNA/cDNA heteroduplexes.
https://www.qiagen.com/us/shop/sample-technologies/dna/dna-clean-up/qiaquick-pcr-purification-kit/#orderinginformation
http://mvz.berkeley.edu/egl/inserts/QIAquick_Spin_Handbook.pdf
Thanx, the purification of cDNA amplicons is clear, but I would like to purify RNA/cDNA heteroduplex ...is it possible?
The answer may depend on the length of products you are using. I would say larger fragments which are expected to be stable in the denaturing conditions provided due to GdHCal will be fine.
You duplex in theory is a double stranded nucleic acid, similar to a PCR product. It may be possible if the product is large and stable. do tell us if it works for you, it will be informative.
Tried to purify an 8kb RNA/DNA hybrid using Quiagen PCR purification kit, eluted with EB buffer. Nanodrop measurements indicated presence of nucleic acids and ran it on a gel but could see nothing. Suspecting that it denatured the hybrids. Anyone else got experience with this?
It is possible to purify a 8 kb nicked circular RNA:DNA hybrid using the Quiagen PCR purification kit using the standard protocol.
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