SacB selection

sacB selection Protocol

For pFastbac-dual or pFastbac vector

1. Digest protein kinase in pDNR-Daul vector with Sal1-HindIII.

2. Digest pFastbac-dual or pFastbac with Sal1-HindIII.

3. Gel purify insert (kinase), ligate to pFastbac-dual or pFastbac vector

4. Transform Top10

5. Grow on LB/Amp/6% Sucrose o/n at 37 oC.

6. Pick single colonies, grow at 37 oC O/N in LB/AMP/6% Sucrose.

7. Make miniprep. Digest with EcoR1-HindIII.