Matt:LabNotes/2014-7-22

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Agi26k0gap Probe Production Continued[edit]

Removal of Amplification Adapters[edit]

To each of 7 x 200ul PCR tubes:

Components Volume
ss-amplicon 71.5
USER 5
H2O 3.5
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