Streptococcus Transformation

Reagents Needed

• CTM (Complete Transformation Medium) pH 6.8, without Calcium Chloride or Bovine Serum Albumin (BSA)
• CTM pH 7.8, with filtered Calcium Chloride added after autoclaving (0.001M CaCl2) and filtered BSA (0.2% BSA)
• CSP-1 peptide (in the -80 freezer) or CSP-2 peptide (which we do not have). Talk to Eric if you are not working with a D39 derivative.

1. Freshly grow up single colonies on a blood TSA plate of the strain to be transformed.
2. Select one colony and grow in 3ml CTM pH 6.8 until OD 0.3, which is 0.39 Absorbance
3. Preheat a microcentrifuge tube of 270ul CTM pH 7.8 to 30 degrees using the hot block.
4. Add CSP-1 peptide to this tube to 0.1 ug/ml eventual final concentration. This is then adding 0.03ug of CSP-1, as our final volume will eventually be 300ul.
5. Add DNA
6. Add 30ul of grown cells (a 1:10 dilution).
7. Vortex
8. Incubate at 30 degrees using the hot block for 30 minutes.
9. Plate cells on a blood TSA plate that has appropriate antibiotics in it.