Week 1

This week we finally started the most important part of our project: the lab work. So of course we had first of all to setup the lab for our needs. Providing schottbottels, flasks, test glasses, ... This week also the first batch of primers arrived so we started the first PCRs.

The PCR for pnikA was done using the Phusion polymerase, the primers pnikA-E,N,X-for and pnikA-S-rev. The annealing temperature was set for 50°C. The template came from the gDNA of the Escherichia coli strain MG1655. (For more information abour the pnikA-system click here).

and prchA

Week 2

Still doing PCRs and start restriction digests.

Week 3

First Biobrick is ready!

Week 4

Week 5

Week 6

Week 7

Week 8

After weeks of hard work the last week of our lab work started. As we already prepared our BioBricks for sending them last week there was only the shipment left. So on Monday we said Good-bye and hope that they were going to have a nice journey to Boston. So until Wednesday, the Wiki-freeze, are only a few tasks left: finishing the testing of our BioBricks BBa_K549001 and BBa_K549002 and of course to prepare the last thins for the european jamboree in Amsterdam. Making the presentation, the poster and of course ordering our team-sweaters.

Protocols

Primer

pnikA-E,N,X-fwd	GCAGAATTCGCGGCCGCTTCTAGAGTTAAGCCTTGCGATCTGCACC
pnikA-S-rev	CCGCTACTAGTAGACGATAAAAGACGCACAAGCC
prcnA-E,N,X-fwd	GCAGAATTCGCGGCCGCTTCTAGAGacggattgtatgagacatggca
prcnA-S-rev	CCGCTACTAGTAcgcaccaagtaagatggcg
luxCDrev	CTGCAGCGGCCGCTACTAGTATTAAGACAGAGAAATTGCTTGAT
PbrR-M,R-fwd	GCAGAATTCGCGGCCGCTTCTAGAGAAGAAGGAGATATACCATGAATATCCAGATCGGCGAG
PbrR-S,A-rev	AGCCTGCAGCGGCCGCTACTAGTAttaCTAGTCGCTTGGATGGGCG
pbrRT-S,A-rev	agtcactagtattaaccggttaTTACACCTGGGTAGATGGCC
pbRMut-fwd	tcgtgcgggattctccagggactgtcggactgc
pbrMut-rev	tccgacagtccctggagaatcccgcacgattgggc
ppbrA-E,N,X-fwd	AGCCTGCAGCGGCCGCTACTAGTAGGTTGCGCGTCGCAACGGAAGC
ppbrA-S-rev	GCAGAATTCGCGGCCGCTTCTAGAGCATGCGGTGCGCTTGGCAAGC
luxCDfor	GAATTCCGCGGCCGCTTCTAGATGGAAAATGAATCAAAATA
luxBfor	ATGAAATTTGGATTGTATGAAATTTGGATTGT
luxBrev	CTGCAGCGGCCGCTACTAGTATTAGGTATATTCC
luxEfor	GAATTCCGCGGCCGcttctagaATGTGACTGGGGTGAGTGA
luxErev	CTGCAGCGGCCGCTACTAGTACTATCAAACGCTTCGGTTAA
iscSfor	AgtcgccggcAAGAAGGAGATATACCATGTACGGAGTTTATAGAGC
icsSrev	agtcactagtattaaccggtctattaatgatgagcccattcg
pabAfor	AgtcgccggcAAGAAGGAGATATACCATGAAATTGCTATTAATTGATAATTATG
pabArev	agtcactagtattaaccggtTTATTACACCACTTTCAAAAAATTATTTAAC
aurFfor	AgtcgccggcAAGAAGGAGATATACCATGCCACGACACCGCGGGC
aurFrev	agtcactagtattaaccggttaTCAACGCGGCGTGTGGGGCG
ChrBAfor	AgtcgccggcAAGAAGGAGATATACCATGAACGCTCTCCCATCCTC
ChrBArev	agtcactagtattaaccggttaTCAGTGATGCAACAACGGATAGG
melAfor	gatcTCTAGAtgGCCGGCGCGTGGCTGGTCGGCAAGCCG
melArev	gatcACCGGTGGCGGACACTATGGCTATTTCTAGC

Team:LMU-Munich/Lab Notebook

From 2011.igem.org