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Old 10-29-2013, 12:41 PM   #1
diniu
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Location: Germany

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Default tRNA sequencing

Hi,
I was wondering whether anybody has ever done with tRNA sequencing from plants(Arabidopsis). I would like to know how to prepare the tRNA
and including precusor tRNA? I would like to purify tRNA from PAA gel, is that ok? How large RNA (70-500nt?) can be purified from the PAA gel?
I would be grateful for any information!
I really need your help!
thank you very much!
Dinu
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Old 10-31-2013, 06:43 PM   #2
HSV-1
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I have done some analyses.
Eventually we didn't go further. There are many rare bases (many are modified ) on tRNA.
These definitely are problems for reverse transcription and therefore for library construction.
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Old 10-31-2013, 11:07 PM   #3
diniu
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Thank you very much for your reply. Yes, modification and secondary structure of tRNA could affect the reverse transcription. But from the literature, it could be done, for example, this paper,6250–6258 Nucleic Acids Research, 2013, Vol. 41, No. 12. I do not know how they did.
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