Team:Groningen/9 September 2010


Assembly of biobricks in pSB1C3-backbone for submission to parts registry by Maarten

O/N cultures of transformants were mini prepped with Sigma Aldrich mini prep kit and control restrictions were performed.

Control restrictions with EcoRI and PstI on constructs:

2 µL construct
2 µL Fast Digest Buffer
0,5 µL EcoRI Fast Digest
0,5 µL PstI Fast Digest
15 µL MQ
Construct Expected band sizes (bp)
pSB1C3-C 2031, 848
pSB1C3-DC 2031, 872
pSB1C3-E 2031, 317
pSB1C3-H 2031, 302
pSB1C3-Sortase 2031, 689
pSB1C3-RFP 2031, 1069


1-3 4-6 7-9
Chp C Chp DC Chp E


1 & 2 4-6 8
Chp H Sortase RFP

The gels were not very conclusive but samples in lane 3, 5 and 7 in the first gel and lane 1 and 6 in the second gel were sent for sequencing on 15 September.

Chaplin detection
Samples from the dry freezer were prepared using TFA.
The samples were diluted with 1 mL Q-H20 and put on gels:
Gel 1: chppt=e deltapt=e bpte e1pte enpte chste deltaste bste e1ste
Gel 2: enste deltapt=4 bpt=4 e1pt=4 enpt=4 deltast=4 bst=4 ent=4
Gel 3: est=4 deltadt=e bdt=e e1dt=e endt=e 1 2 3
Gel 4: 4 e1dt=e bdt=e e1nt=e e1pt=e bpt=e e1st=e b1st=e