Team:UNITN-Trento/Notebook/Labposts/06/67
From 2013.igem.org
{ "date" : "2013-06-27", "author" : "gabriele-emil", "title" : "SAMsynthetase and R0010, the neverending story", "content" : "First of all Gabriele quantified the four pSB1C3+R0010 inocula of yesterday.
Quantification
Sample | Quantity (ng/µl) |
---|---|
#A | 243.7 |
#B | 194.1 |
#C | 245.3 |
#D | 252.9 |
Then, the four quantified samples were screened after a digestion with ExoRI-HF and PstI-HF, with the usual screening protocol (incubated only for 45min, since both the enzymes are HF).
The digestion products were then run on a gel with a transparent loading dye (30% glycerol): each loaded sample contained 16µl of the sample and 4µl of transparent loading dye. Both a 1kb normal ladder and a 100bp transparent ladder were loaded, also the first well was loaded with the usual loading dye.
Gel
Loading scheme |
---|
Loading dye |
Empty |
100bp ladder |
#A |
#B |
#C |
#D |
Empty |
Empty |
1kb ladder |
Finally, Emil prepared the inocula of the product of ligation of R0010 and SAMsynthetase (in pSB1A2: 6 inocula of the 1:1 ligation product and 3 of the 1:3 ligation producta).", "tags" : "SAMsynthetase-Plac" }