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.

  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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