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SEPTEMBER: WEEK 4
September, 20th
Screening PCR on 6 colonies picked from I55 was performed. The results were the follow:
Inoculum of I52, I53, I55, I56 and I57 in LB+Amp for MiniPrep of the following day
September, 21st
Miniprep and Nanodrop quantification for:
- I52 : 92,8 ng/ul
- I53 : 118,5 ng/ul
- I55 : 121,1 ng/ul
- I56 : 159,6 ng/ul
- I57 : 103 ng/ul
For I61 the DNA is already available quantified 147,1 ng/ul.
After 3 hour digestion was performed:
| Culture | Kind | Final reaction volume (ul) | DNA (ul) | H20 (ul) | Enzyme 1 (ul) | Enzyme 2 (ul) | Buffer H (ul)
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| I52 | Screening | 25 | 10,8 | 9,7 | 1 EcoRI | 1 XbaI | 2,5
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| I53 | Screening | 25 | 8,5 | 12 | 1 EcoRI | 1 XbaI | 2,5
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| I55 | Screening | 25 | 14 | 6,5 | 1 EcoRI | 1 SpeI | 2,5
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| I56 | Screening | 25 | 10 | 10,5 | 1 XbaI | 1 PstI | 2,5
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| I57 | Screening | 25 | 9,7 | 10,8 | 1 EcoRI | 1 XbaI | 2,5
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| I61 | Screening | 25 | 6,8 | 13,7 | 1 EcoRI | 1 XbaI | 2,5
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Gel run/cut of samples
As you can see only I52, I56, I57 and I61 are positive so gel extraction was performed on this samples.
Tecan Test on self-inducible promoters was performed on prepared samples with a different protocol that is to bring the cultures to a known OD.
September, 22nd
September, 23rd
September, 24th
September, 25th
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