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For cRNA synthesis, H4 neuroglioma cell lines stably expressing GFP or FLAG-FE65 together with myc-Teashirt3 were harvested and total RNA purified using Qiagen RNA easy kits following the manufacturer's protocol.
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Amplicons were cloned according to the manufacturer's protocol in the pGEM-T Easy kit (Promega) and Sanger-sequenced using an ABI sequencer (Applied Biosystems).
Frozen corneal tissues were ground in liquid nitrogen and DNA was isolated using the DNA easy kit (Qiagen, cat # 69504).
To confirm the SBE genotypes, a subset of PCR products was bacterially cloned with the pGEM®-T EASY kit (Promega) according to manufacturer's instructions.
> > RNA was isolated using Qiagen RNA Easy Kit.
A DNA easy kit and RNeasy Micro Kit were procured from Qiagen (Germany).
For qPCR, RNA was isolated using the Qiagen RNA easy kit including the recommended DNAase treatment.
Total-RNA was isolated with a Qiagen RNA-Plus Mini Easy kit (no.
RNA was extracted using Qiagen RNA easy kit following the manufacturer's protocol.
The genes were cloned using the pGEM-T Easy kit (Promega) and sequenced.
Whole-body RNA from these frozen individuals was extracted using an RNA Easy kit (QIAGEN Inc).
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