IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-7-26: Difference between revisions
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The liquid medium consisted of: | The liquid medium consisted of: | ||
*8 µL LB-min | *8 µL LB-min (We used LB-min because we ran out of regular LB) | ||
*3.2 µL Kanamycin | *3.2 µL Kanamycin | ||
==Primer diagnostic gel== | ==Primer diagnostic gel== | ||
Because of the smearing in the gel from [[IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-7-25#Site_Specific_Mutagenesis_Attempt_5|yesterday's mutagenesis attempt]], we suspected that our primers might be contaminated. To investigate this problem, we made new dilutions of our primers from the master | Because of the smearing in the gel from [[IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-7-25#Site_Specific_Mutagenesis_Attempt_5|yesterday's mutagenesis attempt]], we suspected that our primers might be contaminated. To investigate this problem, we made new dilutions of our primers from the master stocks, and ran 4 of them on a gel along with 4 of the old primer mixes. We did the same with our dNTPs. | ||
The gel was 1.5% agarose, run for 45 minutes. Image | The gel was 1.5% agarose, run for 45 minutes. Image to the right. | ||
[[Image:Cyano primer diagnostic 2006-7-25.jpg]] | |||
Our gel did not detect any contamination in our primers. This doesn't mean they're contaminated; PCR can amplify DNA contamination that the gel doesn't pick up. | |||
Notice that there appear to be no bands in lanes 8 and 9 (old and new EcoR1). We will nanodrop our primer mixes to check the DNA concentrations. | |||
Revision as of 13:59, 26 July 2006
Inoculation of LC1PUR1 and LC2PUR1 transformants
Yesterday's transformants grew succesfully. We inoculated 8 liquid cultures, taking 2 colonies from each of the following plates:
- LC1PUR1 3kb #1
- LC1PUR1 400b #2
- LC2PUR1 3kb #1
- LC2PUR1 400b #2
The liquid medium consisted of:
- 8 µL LB-min (We used LB-min because we ran out of regular LB)
- 3.2 µL Kanamycin
Primer diagnostic gel
Because of the smearing in the gel from yesterday's mutagenesis attempt, we suspected that our primers might be contaminated. To investigate this problem, we made new dilutions of our primers from the master stocks, and ran 4 of them on a gel along with 4 of the old primer mixes. We did the same with our dNTPs.
The gel was 1.5% agarose, run for 45 minutes. Image to the right.
Our gel did not detect any contamination in our primers. This doesn't mean they're contaminated; PCR can amplify DNA contamination that the gel doesn't pick up.
Notice that there appear to be no bands in lanes 8 and 9 (old and new EcoR1). We will nanodrop our primer mixes to check the DNA concentrations.
