Matt:LabNotes/2014-6-19: Difference between revisions
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>Mzcai (Created page with "==Check Agi26k0gap Oligos for DpnII Restriction Sites== *Used ProbesWithDpnIISites.pl to identify oligos with extra DpnII site *Oligos iden...") |
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==Check Agi26k0gap Oligos for DpnII Restriction Sites== | ==Check Agi26k0gap Oligos for DpnII Restriction Sites== | ||
*Lambda exonuclease digestion has been having low efficiency [[Matt:LabNotes/2014-5-22#Probe_Expansion_PCR | Try 1]] and [[Matt:LabNotes/2014-6-11#Lambda_Exonuclease_Digestion | Try 2]] | |||
**Dr. Zhang suggested skipping lambda if very few oligos would be digested | |||
*Used [[File:ProbesWithDpnIISites.txt | ProbesWithDpnIISites.pl]] to identify oligos with extra DpnII site | *Used [[File:ProbesWithDpnIISites.txt | ProbesWithDpnIISites.pl]] to identify oligos with extra DpnII site | ||
*Oligos identified by line # in the [[File:ProbesWithDpnII.txt | text output]] | *Oligos identified by line # in the [[File:ProbesWithDpnII.txt | text output]] | ||
*Of 12,964 oligos, 8,744 (67%) have DpnII restriction sites besides the intended cleavage site | *Of 12,964 oligos, 8,744 (67%) have DpnII restriction sites besides the intended cleavage site | ||
**'''Conclusion:''' cannot skip lambda digestion step |
Latest revision as of 01:40, 20 June 2014
Check Agi26k0gap Oligos for DpnII Restriction Sites[edit]
- Lambda exonuclease digestion has been having low efficiency Try 1 and Try 2
- Dr. Zhang suggested skipping lambda if very few oligos would be digested
- Used ProbesWithDpnIISites.pl to identify oligos with extra DpnII site
- Oligos identified by line # in the text output
- Of 12,964 oligos, 8,744 (67%) have DpnII restriction sites besides the intended cleavage site
- Conclusion: cannot skip lambda digestion step