Cong T. Trinh:heat shock

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Heat Shock

1.Pre-chill 15mL sterile culture tube on ice.

2.Place Competent Cells on wet ice until just thawed.

3.Gently transfer 50 µl of Competent Cells to pre-chilled 15mL culture tube.

4.Add 1-2 µl of Ligated DNA (either Registry BioBricks from distribution kit or ligated parts). Move pipette tip through the cells while dispensing. Gently (VERY GENTLY) tap the tube once or twice.

5.Immediately return tube to ice for 30 min.

6.Place tube in 42 C water bath for 45-50 sec (DO NOT SHAKE).

7.Immediately place the tube on ice for 2 min.

8.Add 950 µl of cold SOC medium and incubate for 60-90 minutes at 37C with shaking.

9.Centrifuge tubes @4000rpm for 3 minutes and discard supernatant. Resuspend cells in remaining solution and use 50μL to spread onto LB with appropriate antibiotic).

10.Incubate the plates overnight at 37C.

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