Bio-rad Ligation and Transformation Module Bedienungsanleitung PDF herunterladen (Seite 39)

b. Prepare starter culture: As late as possible the day before the

transformation, inoculate a 2–5 ml LB culture with a starter colony

from the

E. coli

starter plate. Incubate cultures in a shaking water

bath or incubator overnight at 37°C and at least 200 rpm.

Note: It is important to use a fresh starter culture (<24 hours since

inoculation) for the transformation, or transformation efficiency will be

reduced.

Note: If a shaking water bath or incubator is not available, incubate starter

cultures at 37°C 1 day prior to the transformation, manually shaking the

vial containing the culture as frequently as possible to oxygenate the

culture (a reduction in transformation efficiency may be observed).

3. Place 1 LB Amp IPTG agar plate per student team in 37°C incubator

just before the transformation lab. Note: If performing a positive control

transformation, a second plate per student team will be needed.

4. Pipet 1.5 ml of C-growth medium to one 15 ml culture tube per team

and incubate at 37°C for at least 10 min prior to the transformation

laboratory.

1 2 ... 34 35 36 37 38 39 40 41 42 43 44 ... 59 60

Keine Kommentare

Bio-rad Ligation and Transformation Module Bedienungsanleitung Seite 39