208 lab

Main purposes today

• helloworld-project
• OD meassurements
• autoclaving
• make competent cells

Who was in the lab

Kristian, Jakob, Henrike

Procedure

OD measurements

placed on ice

Making competent cells

Using the Protocol, we got from Andreas with following modifications to protocol:

• made only half the amount
• used falcon tubes

The competent cells were stored in -80 C freezer on the lower shelf

Autoclaving

following was autoclaved:

• Eppendorf tubes
• 3 x 100ml dm water
• 2 x 100ml 0.1M CaCl_2 and 15% glycerol
• 1 L bottle of 0.1M CaCl_2
• 1 L bottle of 50% glycerol
• 200ml bottle of 1M CaCl_2

Solutions made

• 0.1 M CaCl_2 and 15% glycerol from 10ml 1M CaCl_2
• 30ml 50% glycerol 70ml dm water
• 0.1 M CaCl_2 from 100ml 1M CaCl_2 and 900ml dm water
• chlorampenicol stock 1ml with cencentration 50mg/ml

Plates

• 24 ps with 10ug/ml chlorampenicol (CAP) 5 mg to 500ml
• 33 ps with 30ug/ml Kanamycin 0.3ml of 30ug/ml to 500ml

Transformation

Following the [http://parts.igem.org/Help:Protocols/Transformation official iGEM protocol] :

• BBa-I0500 plate 5 well 14N, then moved to -80C
• BBa-I14032 plate 3 well 18H, then moved to -80C

Transformation efficiency test (0.5,5,10,20,50)

Incubation 2 hours with 300 rpm

Plates:

• I0500 - no incubation on Kanamycin plate, one 20ul and one 200 ul
• I14032 - no incubation on CAP, one 20ul and one 200 ul

all 4 had 2 hours incubation.

Navigate to the Previous or the Next Entry