Noi/NOTES/2012-6-5
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Randomly tagging primer experiment
Sanger sequencing results of the amplicons amplified with randomly tagging primers (with USER)
Check Sanger's sequencing result roughly before sending more clones
- Primimg with Syb_RP-7: read reverse complementary of the AmpFA/NU.Sol strand
qs (quality score) Sequence correct 7nt=AAAAAAA Note contain AGAGTG(7)GTG 1U-1 29 seem to (overlapping peaks) yes, :Homopolymeric or Repetitive Region, **request for free repeat 1U-2 29 yes yes, :Homopolymeric or Repetitive Region 1U-3 33 yes yes, chromatogram very clear even the peak very low 1U-4 32 yes yes, 1U-5 42 yes yes, 1U-6 43 yes yes, 1U-7 42 yes yes, chromatogram very clear even the peak very low 1U-8 14 seem to (overlapping peaks) yes Non-specific, **request for free repeat 1U-9 31 yes yes, 1U-10 43 no no, This clone has a shift band higher than other positive clones on E-gel will look closer to the sequences (seem to be neither AmpFNU.Sol nor AmpFAU.Sol)
- Summary: From 10 clones sequences by Sanger's sequencing
- 7 clones are clearly correct and all contain 7T
- 2 clones clearly showed 7T but show overlapping of the peaks surrounding 7T sequences --> request for free repeat
- 1 clone showed unrelated sequences of the clone amplified by AmpFNU.Sol or AmpFAU.Sol
- Images of trace file will be uploaded later