Digestion of PMP and PGL4.30 for ligation

prepared PMP2 , PGL2 and PGL3 for ligation

restriction enzyme digest mixtures were as follows:

PMP2 control for plasmid

DNA 2µL HindIII 1µL XbaI 1µL N4 buffer 10x 5µL BSA 100x 0.5µL Water 40.5 µL

PMP2 control for NotI

DNA 2µL NotI 1µL N4 buffer 10x 5µL BSA 100x 0.5µL Water 41.5 µL

PMP2 control for SpeI

DNA 2µL SpeI 1µL N4 buffer 10x 5µL BSA 100x 0.5µL Water 41.5 µL

All let 1:30 hours at 37ºC, shaking (600 rpm).

Bands got in the control of DNA and enzymes. Everything seems to be correct now!

After we ran the digestion products in a gel (1% agarose, 50 µl of digestion product + 10 µl of loading dye).

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