Team:Hawaii/Notebook/2008-07-21

= Things we did today =

Colony PCR



 * Margaret


 * Colony PCR of pSB1A2, pSB1A3, pSB3K3

Results


 * Lane 2 contains bands that are approximately 1kb, while I would expect pSB1A3 to be 991bp. Lane 3 containing pSB1A2 should be 913bp and is just above the 0.9kb mark. Lane 4 contains pSB3K3 and should be 991bp and is just below 1kb.
 * The positive control, UPA amplifying algal DNA is at 0.5kb as expected.
 * The negative control, UPA amplifying water, contains no band.
 * In lane 7 pSMC121, a plasmid containing the omega interposon was run, but no DNA was stained.

Conclusions


 * The plasmids (pSB1A3, pSB1A2, pSB3K3) were verified using VF2 and VR. I will make a glycerol stock and do a plasmid prep.


 * The absence of a a band for pSMC121 may explain the difficulty I have had in amplifying the omega region of this plasmid. I need to go get a new sample.

PCR amplification of pRL1383a parts

 * continued from Saturday, ran a gel

Restriction Digest

 * Krystle
 * Performed 2 restriction digests on nir, gfp, and gfp fusion brick with EcoRI and SpeI
 * trial 1: digested 5ul of each plasmid prep in a 50ul reaction mixture, ran 20 ul of the digest after 3 hours
 * trial 2: digested 25ul of each plasmid prep in a 50ul reaction mixture, ran 20 ul of the digest after 2 hours

Name of Task

 * Krystle, Margaret


 * We figured out why some of the biobrick extractions did not transform. Some of these parts are on pSB2K3[] which is a low copy until IPTG is added to the medium in which case it becomes high copy! So next time we make this, add 1mM IPTG to the media.

= Discussion =

= Quote of the Day = "History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson"