"

From 2013.igem.org

Just testing this right now...

Experiment 1.1 – Chemical Transformation Efficiency

Purpose:

To explore the effects of competent cell : DNA ratio and heat shock temperature and duration on chemical transformation efficiency.

Setup:

Trial V (μL) cells V (μL) pUC19 Heat Shock Resistance 1 (2x) 50 0 - - 2 (2x) 170 1 10 min @ 37° C Ampicillin 3 (2x) 50 2 10 min @ 37° C Ampicillin 4 (2x) 50 2 30 s @ 42° C Ampicillin 5 (2x) 50 2 60 s @ 42° C Ampicillin

Procedure: 1. Thaw competent cells on ice for ~15 minutes 2. Aliquot 50 mL competent cells into 1.5 mL Eppendorf tubes 3. Add pUC19 into competent cell aliquots 4. Incubate on ice for ~30 minutes 5. Use water bath to heat shock samples at various conditions 6. Add 1 mL SOC recovery medium to each sample 7. Allow cells to recover in shaker @ 37° C for ~1 hr. 8. Plate 200 μL of each sample and allow to dry 9. Store plates upside down in incubator @ 37° C overnight Results: Discussion: