Team:SDU-Denmark/labwork/Notebook/week9

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iGEM TEAM ::: SDU-DENMARK

Laboratory Notebook

1st week 2nd week 3rd week 4th week
5th week 6th week 7th week 8th week
9th week 10th week 11th week 12th week

27-08-2012 to 02-09-2012

DNA purification from FFT liquid cultures were made.

Nanodrop was performed on it:
1: 83,6 ηg/μL
2: 17,2 ηg/μL
3: 121,1 ηg/μL
4: 48,1 ηg/μL
5: 99,1 ηg/μL
6: 59,6 ηg/μL
7: nothing(an error must have occurred)
8: 59,6 ηg/μL

We made check-digest with EcoRI+SpeI to examine the plasmids in preparation of mutagenesis. The check digest showed that some had the appropriate digested sizes we would expect from a plasmid of 3kb and a gene of 2kb.

Mutagenesis was run on these FFT colonies containing all primers.
We made new SST cultures again and plated them on AMP-resistance plates.
Isolated FFT and SST plasmids and sent them for sequencing.
Afterwards we did a digest and transformation on FFT and plated them.
We made liquid cultures of FFT and SST and incubated them over night. Later we made transformation on the three parts we chose from the iGEM kit.
1) R0010
2) E0040
3) B0015
Afterwards we plated them on AMP-resistance plates.
We made liquid cultures of the three parts

Purification of SST, FFT, B0015, E0040 and R0010 and following PCR


The O.N. cultures of SST, FFT and our 3 parts (B0015/terminator, E0040/GFP and R0010/LacL) showed promising results of growth. A miniprep was made from the 32 samples and a Nanodrop as well which gave the following results:

NanoDrop Results
1. R0010, culture 1 18,0 ng/ul2. B0015, culture 3 67,9 ng/ul
3. B0015, , culture 562,9 ng/ul4. R0010, culture 3 70,5 ng/ul