Team:UNITN-Trento/Notebook/Labposts/06/66

From 2013.igem.org

{ "date" : "2013-06-29", "author" : "emil", "title" : "Purification and screening of the GFPs E0840 and E0240", "content" : " I did the miniprep of the 8 inocula(2 GFP constructs) following the vacuum protocol then I have quantified the products with the following results:

Quantification
SampleQuantification
BBa_E0840/1 484.6ng/µl
BBa_E0840/2 376ng/µl
BBa_E0840/3 481.8ng/µl
BBa_E0840/4 475ng/µl
BBa_E0240/1 361ng/µl
BBa_E0240/2 454.4ng/µl
BBa_E0240/3 368ng/µl
BBa_E0240/4 365.8ng/µl
Then I did the screening of the samples E0840/1,E0840/3,E0240/2,E0240/4. I digested 800 ng of each sample with EcoR1-HF and Pst1-HF following the digestion protocol for screening. After 40 minutes I loaded the samples on a gel with the following results:
Gel order
SampleWell
Ladder 1kb Fermentas1
BBa_E0840/12
BBa_E0840/33
BBa_E0240/24
BBa_E0240/45
We can see 2 bands for well: the upper is the backbone(pSB1A2 2079 bp), the lower is presumably the GFP(BBa_E0240 826 bp,BBa_E0840 828 bp)(pefix and suffix escluded ?).", "tags" : "E0840-E0240" }