When I used 95bp PCR product (full-length tRNA with promoter), the T7 polymerase RNA synthesis kit (NEB) gave a lot of large bands (more than 200bp). Can you explain the reason and the way to remove all the bigger RNA bands.
Yun-Tzai Lee
First, you need to confirm that apart from the target band, these is no any other bands observed in your PCR product which is larger than 95 bps. If the PCR product is pure, you may need to purify the tRNA transcripts using denaturing gel and verify its molecular weight by Mass spectrometry after purification.
Good luck.
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