Search found 9 matches

by javaid
Fri Jul 13, 2007 1:00 pm
Forum: Molecular Biology
Topic: plasmid isolation
Replies: 0
Views: 1056

plasmid isolation

im using qiagen giga kit to isolate my plasmid but am this time short of filter cartriges, is there any alternative to that.... can i use millipore filtration unit instead, havig a .22 micron membrane
by javaid
Fri Apr 20, 2007 1:00 pm
Forum: Molecular Biology
Topic: smear after restriction digestion
Replies: 23
Views: 32615

u may try by incubating the sample without buffer...if it gets smeared then the problem is with enzyme...as per u both the mini n midi prep had the same results then it is the thing that is common in both isolations n that hopefully is only the enzyme so u may try with other enzyme ...... hope this ...
by javaid
Sun Feb 25, 2007 5:20 pm
Forum: Molecular Biology
Topic: smear after restriction digestion
Replies: 23
Views: 32615

no i does not have star activity whereas my earlier preprations of plasmids give good results after using these enzymes...
by javaid
Sat Feb 24, 2007 7:41 am
Forum: Molecular Biology
Topic: smear after restriction digestion
Replies: 23
Views: 32615

actually it gets smeared only during restriction digestion coz when i run uncut sample it gives good bands but gets smeared after restriction digestion n even if i dont add the enzyme n incubate only with buffer... so i think it is nucleases which get active in buffer... now i need to purify my samp...
by javaid
Fri Feb 23, 2007 4:07 pm
Forum: Molecular Biology
Topic: smear after restriction digestion
Replies: 23
Views: 32615

actually the problem is that earleir plasmid isolations by the same kit give quite good results after digestion but the last two preprations give the smear... so i think it is nuclease contamination ... but now i need to purify that sample n get rid of nucleases from that sample .is it somehow possi...
by javaid
Fri Feb 23, 2007 11:40 am
Forum: Molecular Biology
Topic: smear after restriction digestion
Replies: 23
Views: 32615

i isolated my plasmid by sigma mini prep kit. uncut sample runs fine and gives 3 bands but once i digest it with enzyme it gives smear more just icubating with buffer n without adding any enzyme it also leads to smear,, i think there is somewhere nuclease cintamination .. can anyone help where it co...
by javaid
Thu Feb 22, 2007 8:43 am
Forum: Molecular Biology
Topic: plasmid prcipitation
Replies: 6
Views: 4264

[quote="LilKim"]
but i think adding salt will enhance the precipitation.. will it ppt without salt.. n if not then which salt to use n conc. thanx
by javaid
Thu Feb 22, 2007 5:02 am
Forum: Molecular Biology
Topic: smear after restriction digestion
Replies: 23
Views: 32615

smear after restriction digestion

my plasmid prepration gives a total smear following restriction digestion.. what may be the possible reason.... need help
by javaid
Thu Feb 22, 2007 4:58 am
Forum: Molecular Biology
Topic: plasmid prcipitation
Replies: 6
Views: 4264

plasmid prcipitation

i have isolated plasmid and is finally suspended in TE but i need it be dissolved in PBS coz i need it for transfection, so may i know how to precipitate the plasmid. do i need to add salt...