lab 3 note 4/21/09

the following steps are after PCR product have been purify with column

1. perform a ligation reactions (this was done by DxRR)
2. add 1 ul of purify PCR product to tube
3. incubate for 5 mins in 70 degree C water bath, then cool on ice for > 2 mins
4. centrifuge, add T4 DNA ligase and pTet plasmid to tubes (again with the help of DxRR)
5. centrifuge, incubate for 10 mins at 37 degree C
6. incubate on ice for > 5 mins

a transformation test was performed by following the transformation handout from DxRR.
except:

  1. 16) use all 10 ul
  2. 17) use all 120 ul of competent cell
  3. 19) plate 60 ul on each LB and on LB + Amp + IPTG
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