Team:LMU-Munich/Lab Notebook

From 2011.igem.org

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(Week 1)
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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.  
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.  
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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).  
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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. ([https://2011.igem.org/Team:LMU-Munich/Project/Description For more information about the pnikA-system click here]).  
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and prchA
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The PCR for prcnA was done using the Phusion polymerase, the primers prcnA-E,N,X-for and prcnA-S-rev. Conditions for the Annealing temperatur were at 50°C. The template was again the gDNA of the E.coli strain MG1655. ([https://2011.igem.org/Team:LMU-Munich/Project/Description For more information about the prcnA-system click here]).
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Revision as of 12:21, 20 September 2011


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 about the pnikA-system click here).

The PCR for prcnA was done using the Phusion polymerase, the primers prcnA-E,N,X-for and prcnA-S-rev. Conditions for the Annealing temperatur were at 50°C. The template was again the gDNA of the E.coli strain MG1655. (For more information about the prcnA-system click here).

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