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EPF-Lausanne/6 October 2009
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==Wet Lab== | ==Wet Lab== | ||
| + | ===2nd gel=== | ||
| + | We did a second gel of yesterday's colony PCR to check the gel as iti is strange not to have any double transformant but we had double antibiotic selection. | ||
| + | ===New colony PCR=== | ||
| + | We think that maybe if the first colony PCR didn't show any double transformants it is because we didn't let enough time for the extansion (1'20'') and RO2 is 1900 bp long and Taq only goes at 1 kb / minute. From now, we will let an extansion time up to 2 minutes. | ||
| + | |||
| + | ===Replating=== | ||
| + | ... of RO2.4 + BB1 (3 Trp K.O. strains), RO1.1 + BB1 (3 strains). | ||
| + | |||
| + | ===Gel=== | ||
| + | Still no double transformants !!! | ||
| + | |||
| + | ===Culture=== | ||
Prepared cultures of 2 x 7mL of DH5 alpha RO2.4 + BB1 clone n.3 (the strain that works) in LB for tomorrow's experiment (the cells were taken from the glycerol stock). Left cells to incubate overnight at 37°C. | Prepared cultures of 2 x 7mL of DH5 alpha RO2.4 + BB1 clone n.3 (the strain that works) in LB for tomorrow's experiment (the cells were taken from the glycerol stock). Left cells to incubate overnight at 37°C. | ||
Latest revision as of 07:12, 8 October 2009
Contents |
Wet Lab
2nd gel
We did a second gel of yesterday's colony PCR to check the gel as iti is strange not to have any double transformant but we had double antibiotic selection.
New colony PCR
We think that maybe if the first colony PCR didn't show any double transformants it is because we didn't let enough time for the extansion (1'20) and RO2 is 1900 bp long and Taq only goes at 1 kb / minute. From now, we will let an extansion time up to 2 minutes.
Replating
... of RO2.4 + BB1 (3 Trp K.O. strains), RO1.1 + BB1 (3 strains).
Gel
Still no double transformants !!!
Culture
Prepared cultures of 2 x 7mL of DH5 alpha RO2.4 + BB1 clone n.3 (the strain that works) in LB for tomorrow's experiment (the cells were taken from the glycerol stock). Left cells to incubate overnight at 37°C.
People in the lab
Gab, Christian
