Team:Groningen/9 September 2010


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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



The gels were not very conclusive but samples 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