Streptococcus suis Transformation

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Day 1:

  • Plate strains from freezer onto Tryptic Soy Plates for single colonies.

Day 2:

  • Put 3ml of Todd-Hewitt + 0.5% Yeast Extract into sterile 16mm test tubes. Add a single colony into the tube; grow overnight at 37 degrees in the plate incubator (so, without shaking).

Day 3:

  1. Put 5ml of Todd-Hewitt + 0.5% Yeast Extract into sterile 16mm test tubes to pre-warm the media for 1 hour.
  2. Add 125 ul of the overnight growth (a 1:40 ratio).
  3. Allow to grow to an Absorbance of 0.035 - 0.058 (one hour, generally).
  4. Put 100ul of this culture into a micro-centrifuge tube.
  5. Add in the correct XIP peptide from the stock to create a final concentration of 250uM. (normally, 7-10ul of stock peptide)
  6. Add 0.5pMol of DNA (1.4ug for pACYC194; see
  7. Vortex; place in 37 degree incubator for 2 hours.
  8. Plate onto appropriate antibiotic Tryptic Soy plates.