Team:Brown/Notebook/June23

From 2010.igem.org

Wednesday, June 23 2010

After incubation for ~16 hours at 37°C, plates BL21 20ng and 100ng and XL1 20ng and 100ng, all the plates seemed to have too many colonies. This may be due to our amp+ plates being too old (degraded amp) so we are growing a single colony in a culture tube (LB + amp; 100µg/ml) at 37°C again.

Picked two colonies each:

  • BL21 20 ng
  • BL21 100 ng
  • XL1 20 ng
  • XL1 100 ng

BL21 samples turned cloudy after ~5 hour incubation; transformation likely worked.

XL1 samples showed no growth in amp+ LB; transformation probably did not work.

Transformation Protocol

  1. Adrian had plasmid at ~150 ng/µl diluted to 10 ng/µl
  2. Add plasmid to 150 µl of cells; low concentration = 20 ng; high = 100 ng of plasmid
  3. Add LB until final volume = 300 µl
  4. Streaked plates (Amp) from BIOL1220 preparation; incubate overnight (6:00 PM to 10:00 AM next day)

In morning: all 4 varieties (BL21 hi, low; XLI high, low) showed extensive colony formation on amp+ plates (possible amp was faulty?)

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