Team:Heidelberg/Templates/Del week17 overview

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Validation of pFSN

We screened for more colonies with pFSN but we didn't find any more positive ones.

Furthermore we did some tests for the validation of pFSN in colony D8w.

Validation of pFSN in clone D8w
Method Details method Result
Screening for inserted fragments Fragments pSB4K5 and DelA-F
Heidelberg Yes check.svg.png
Fragments DelA-F and DelF-G
Heidelberg Yes check.svg.png
Restriction Digest EcoRI
Heidelberg Yes check.svg.png
BamHI
Heidelberg Yes check.svg.png
BglII
Heidelberg Yes check.svg.png
Sequencing gibson overlaps Primer FS_13_short
Heidelberg Yes check.svg.png
Primer FS_22
Heidelberg Yes check.svg.png
Primer VFII
Heidelberg Yes check.svg.png
Primer VR
Heidelberg Tilde.png

There is a mutation at the beginning of the mRFP cassette. This explains why the colonies do not get red. All other tests for valiation were positive. Consequently colony D8w includes the right plasmid. Because we cannot use mRFP as expression control we will do SDS-PAGE instead to prove that the proteins are expressed.