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After treatment by DNaseI (DNA-free TM Kit, Ambion), purified RNA was proceed with reverse transcription using the Super Script III First-strand kit (Invitrogen) to obtain first-strand cDNA.
In-gel digestion was proceed with 0.01 mg trypsin (Sigma Aldrich) over night at 37°C.1 µl of the digested peptide was spotted on a Anchorchip treated with matrix solution CHCA(Sigma) and washing with 0.1% TFA, then the crystallized sample was subjected to MALD-TOF-MS (ABI4700).
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com