Matt:LabNotes/2014-7-22: Difference between revisions

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>Mzcai
(Created page with "==Agi26k0gap Probe Production Continued== ===Removal of Amplification Adapters=== *[[Matt:LabNotes/2014-7-1#Lambda_Exonuclease_Digestion | Lambda digested 14.2ug (200ul) ssDNA...")
 
>Mzcai
mNo edit summary
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*[[Matt:LabNotes/2014-7-1#Lambda_Exonuclease_Digestion | Lambda digested 14.2ug (200ul) ssDNA]]  
*[[Matt:LabNotes/2014-7-1#Lambda_Exonuclease_Digestion | Lambda digested 14.2ug (200ul) ssDNA]]  
*[[Matt:LabNotes/2014-7-17#Lambda_Exonuclease_Digestion | Lambda digested 21.4ug (320ul)ssDNA]]
*[[Matt:LabNotes/2014-7-17#Lambda_Exonuclease_Digestion | Lambda digested 21.4ug (320ul)ssDNA]]
*Combine to 300ul of 22ug total ssDNA
*Combine to 520ul of 35ug total ssDNA


To each of 7 x 200ul PCR tubes:
To each of 7 x 200ul PCR tubes:

Revision as of 18:24, 22 July 2014

Agi26k0gap Probe Production Continued

Removal of Amplification Adapters

To each of 7 x 200ul PCR tubes:

Components Volume
ss-amplicon (73.3 ng/ul) 75
USER 5
H2O 0
total 80
 *5ug ssDNA per tube
  • Incubated at 37C for 2 hours
  • Added 15ul of the following
Components Volume
10X DpnII Buffer 10
100uM RE-DpnII guide oligo 5
 RE-DpnII guide oligo:
 0-gap: RE DpnII V6
  • Incubated at 94C for 2 min, then 37C for 3 min
  • Added 5ul DpnII
  • Incubated at 37C for 3 hr
  • Added 100ul of the following
Components Volume
10X DpnII Buffer 10
USER 5
H2O 85
Total 100
  • Incubated at 37C for overnight (~15 hrs)

Summary: 7 tubes of 200ul each