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Team:SDU-Denmark/labwork/Notebook/week9
From 2012.igem.org
Laboratory Notebook
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:
| 1. R0010, culture 1 | 18,0 ng/ul | 2. B0015, culture 3 | 67,9 ng/ul |
|---|---|---|---|
| 3. B0015, , culture 5 | 62,9 ng/ul | 4. R0010, culture 3 | 70,5 ng/ul |
