Glasgow/Wetlab/Week9

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Gels

Week 9

Maija performed a PvuI digest for pDntA+3/20G and pDntA+4/6B constructs with Roche's PvuI enzyme. The bands were weird sizes, see the gel here.

Insert Vector Enzyme Expected size Result

pDntA 3/20G PvuI 1450bp + 749bp ~2500bp

pDntA 4/6B PvuI 2375bp + 681bp ~3800bp + ~1250bp
Christine digested minipreps of interest B, C and D suspected to contain (*d→g*) from 24/08/07 with PstI. Miniprep of colony D shows bands of expeceted sizes 1155bp, 549bp, ~1962bp, and 3682bp.
Overnights were also set up of colonies K, L, M, N, Q, S and T, each suspected of containing (*a→d*), from plate 2.
Tuesday 28 August 2007

Wednesday 29th August 2007

Scott did colony PCR (see protocol 9) on the assembly transformations carried out yesterday using primers VF2 and VR. Unfortunately it looks as though the combined product is larger than the sum of its parts...
Scott also did the following restriction digests (see Protocol 7):

Insert Digest 1 (EcoRI/PvuI) Insert Digest 2 (EcoRI/SpeI)
(Done overnight at 16°C)

J23119 (tube 1) J61101

J23119 (tube 2) "

Destination (EcoRI/XbaI)
(Done overnight at 16°C)

J61101

Thursday 30th August 2007

Friday 31st August 2007

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Insert	Vector	Enzyme	Expected size	Result
pDntA	3/20G	PvuI	1450bp + 749bp	~2500bp
pDntA	4/6B	PvuI	2375bp + 681bp	~3800bp + ~1250bp

Insert Digest 1 (EcoRI/PvuI)	Insert Digest 2 (EcoRI/SpeI) (Done overnight at 16°C)
J23119 (tube 1)	J61101
J23119 (tube 2)	"