IGEM:metu/2009/Notebook/wound dressing/2009/08/04

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1. 10:00 -- Plates ,which are transformed, were taken from incubation and put into +4 C.

2. 11:30 –- Plasmid isolation of backbone ( 4 tane)

They were code as BB-10, BB11, BB12, BB13

3. 12:00 -- Electrophrose of LUXR-RBS, Term B0010, Term B0012 and RBS because nanodrop results of them were very high.

4. 16:30 -- Digestion of LUXR-RBS

LUXR-RBS     EcorI and SpeI = Buffer R – 4 fold excess of speI ( commercial)
However, we used fast digest. Same buffer is used with EcorI and SpeI in fast digestion process.

1 microgram of DNA was used. 
14 µl water- nuclease free
2 µl buffer
1.63 µl DNA
1 µl EcorI + 1 µl speI
Wait for 1 hour at 37 C.
Results: Only Backbone was cut.

5. Preparation of Agorose gel for ligation.

while loading to gel;
20 µl plasmid + 4 µl loading dye   (no water)			           
5 µl Ladder