pET28b digestion problem - pET28b plasmid with CASC2a sequence (Aug/03/2007 )
hi
i used pET28b plasmid containing CASC2a sequence. i did digestion but on the gel i didnt get the band of digestion of pET28b plasmid. as i run the undigested plasmid along with digested i get the band of undigested but didnt get band of digested. digested was done with nde1, sal 1 seperate and both togather. so what could b the problem with digestion.
best regards animesh patel
-animesh patel-
QUOTE (animesh patel @ Aug 3 2007, 04:54 AM)
hi
i used pET28b plasmid containing CASC2a sequence. i did digestion but on the gel i didnt get the band of digestion of pET28b plasmid. as i run the undigested plasmid along with digested i get the band of undigested but didnt get band of digested. digested was done with nde1, sal 1 seperate and both togather. so what could b the problem with digestion.
best regards animesh patel
i used pET28b plasmid containing CASC2a sequence. i did digestion but on the gel i didnt get the band of digestion of pET28b plasmid. as i run the undigested plasmid along with digested i get the band of undigested but didnt get band of digested. digested was done with nde1, sal 1 seperate and both togather. so what could b the problem with digestion.
best regards animesh patel
Maybe you can change the exposure time of your camera. For example, you enlong 1 second to 10 second, then you maybe see the band.
-susion888-
QUOTE (susion888 @ Aug 3 2007, 05:16 AM)
QUOTE (animesh patel @ Aug 3 2007, 04:54 AM)
hi
i used pET28b plasmid containing CASC2a sequence. i did digestion but on the gel i didnt get the band of digestion of pET28b plasmid. as i run the undigested plasmid along with digested i get the band of undigested but didnt get band of digested. digested was done with nde1, sal 1 seperate and both togather. so what could b the problem with digestion.
best regards animesh patel
i used pET28b plasmid containing CASC2a sequence. i did digestion but on the gel i didnt get the band of digestion of pET28b plasmid. as i run the undigested plasmid along with digested i get the band of undigested but didnt get band of digested. digested was done with nde1, sal 1 seperate and both togather. so what could b the problem with digestion.
best regards animesh patel
Maybe you can change the exposure time of your camera. For example, you enlong 1 second to 10 second, then you maybe see the band.
but i also run the digest of pET33b along with pET28b, band for pET33b digest was visible where as with pET28b wasn't. so i think there should be problem with digestion site in pET28b but cant judge the problem. so can u guide me for that.
thankss
-animesh patel-