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Team:Stockholm/17 August 2010
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Andreas
Site-directed mutagenesis
Continued from 16/8
Glycerol stocks
From 16/8 ON cultures
yC1: pSB1C3.m.yCCS 1, 17/8 yC4: pSB1C3.m.yCCS 4, 17/8 SC1: pSB1C3.mut SOD 1, 17/8 SC2: pSB1C3.mut SOD 2, 17/8
1600 μl cell culture in 400 μl 100 % glycerol
Plasmid prep
| DNA concentrations | ||||
|---|---|---|---|---|
| Prior to concentration | After concentration | |||
| Sample | Conc [ng/μl] | A260/A280 | Conc [ng/μl] | A260/A280 |
| yC1→pSB1C3.m-yCCS 1 | 36.88 | 1.82 | 250.2 | 1.92 |
| yC4→pSB1C3.m-yCCS 4 | 41.83 | 1.77 | 215.7 | 1.87 |
| SC1→pSB1C3.m-SOD 1 | 47.38 | 1.72 | 230.2 | 1.91 |
| SC2→pSB1C3.m-SOD 2 | 38.00 | 1.72 | 104.0 | 1.86 |
From 16/8 ON cultures
Elution volume: 70 μl dH2O
Since samples were intended for sequencing, they had to be concentrated in the evaporator. Results in table.
Samples sent for sequencing from VF2 primer annealing site:
- 15 μl DNA
- 1.5 μl 10 mM VF2 primer
ON cultures
New ON cultures (5 ml + 25 Cm) were set for plasmid prep from same clones as were sent for sequencing: yC1, yC4, SC1, SC2.
