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EPF-Lausanne/25 July 2008
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Our electroporation didn't work :-( No bacteria did grow... | Our electroporation didn't work :-( No bacteria did grow... | ||
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We also received part P0140, but it hasn't grown during the night, we have to do it once more on monday. | We also received part P0140, but it hasn't grown during the night, we have to do it once more on monday. | ||
| - | We did minipreps for these 15 parts | + | We did minipreps for these 15 parts. |
| + | |||
| + | We received our Primers so we did a pcr of 2 parts(one containing RhlR) and 2 of the Ron-Weiss placmids(that contained LuxR). | ||
| + | We used 3 sets of primers. 1: Biobrick primer, 2: RhlR primer, 3: LuxR primer | ||
| + | We coupled every bit of DNA with each primer once to have negative controls. So a total of 12PCRs | ||
| + | Protocol: | ||
| + | prepared mix: | ||
| + | 10x PCR buffer: 2.5ul | ||
| + | MgCl: 0.75ul | ||
| + | dNTP:: 0.5ul | ||
| + | polymarase: 0.1ul | ||
| + | water 19.65 | ||
| + | DNA: 0.5ul | ||
| + | primers: 0.5ul each | ||
Latest revision as of 12:13, 7 August 2008
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Our electroporation didn't work :-( No bacteria did grow...
We did some glycerol stocks from the swipes parts we received yesterday : - pSB4K5 - pSB4C5 - pSB3K3 - pSB1AC3 - pSB1A3 - pSB4A3 - B0034 - R0040 - E1010 - I14033 - F1610 - B0015 - I0466 - C0070 - R0071 They are stored at -80 degrees
We also received part P0140, but it hasn't grown during the night, we have to do it once more on monday.
We did minipreps for these 15 parts.
We received our Primers so we did a pcr of 2 parts(one containing RhlR) and 2 of the Ron-Weiss placmids(that contained LuxR). We used 3 sets of primers. 1: Biobrick primer, 2: RhlR primer, 3: LuxR primer We coupled every bit of DNA with each primer once to have negative controls. So a total of 12PCRs Protocol: prepared mix: 10x PCR buffer: 2.5ul MgCl: 0.75ul dNTP:: 0.5ul polymarase: 0.1ul water 19.65 DNA: 0.5ul primers: 0.5ul each
