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From 2013.igem.org

Validation of pFSN

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

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

Results

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

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.