Team:UNITN-Trento/Notebook/Labposts/06/57

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Revision as of 07:47, 9 September 2013

{ "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

Sample	Quantification
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

Sample	Well
Ladder 1kb Fermentas	1
BBa_E0840/1	2
BBa_E0840/3	3
BBa_E0240/2	4
BBa_E0240/4	5

image

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" }