Matt:LabNotes/2014-6-24: Difference between revisions

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(Created page with "==Removal of Amplificiation Adapters== * Lambda digested 13.7ug ssDNA *[[Matt:LabNotes/2014-5-22#Lambda_Exonuclease_...")
 
>Mzcai
mNo edit summary
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*Incubated at 94C for 2 min, then 37C for 3 min
*Incubated at 94C for 2 min, then 37C for 3 min
*Added 5ul DpnII
*Added 5ul DpnII
*Incubated at 37C for 2 hr
*Incubated at 37C for 3 hr
*Added 100ul of the following
*Added 100ul of the following
{| {{table}}
{| {{table}}

Revision as of 22:58, 24 June 2014

Removal of Amplificiation Adapters

To each of 4 x 200ul PCR tubes:

Components Volume
ss-amplicon (73.3 ng/ul) 75
USER 5
H2O 0
total 80
 *5.5ug 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: 4 tubes of 200ul each