User:Barry Canton/Notebook/T7 RNAP transcription of rRNA/2008/08/21

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Previous entry T7 RNAP transcription of rRNA

Ok, I ran a gel today using the old extracts again, no reason to think they might be bad. I followed the protocol pretty closely. The only things that changed were -

  • I did 6μl of extract so that I could split in two. So I added 30μl of glyoxal reaction mix to each. This means that once I have added 50μl of formamide buffer that I can only add about half of the RNA to the gel.
  • My glyoxalated RNA samples sat on ice for about an hour because I needed to make and autoclave the BTPE.
  • I ran the gel for four hours
  • I only did 20 min washes in the denaturing buffer, neutralizing buffer, and 20xSSC.


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