DNA gel electrophoresis problem? - (Jun/25/2008 )
can DNA heated upto 90C be used for electrophoresis?
if sonication or Rest. Enz. are not available, is there a way to fragment DNA and run them for viewing by gel staining?
i tried but couldnt get proper bands as i was told its because os denaturation of DNA.....
will heat treated dna wont give bands???
please help
[quote name='frost' date='Jun 25 2008, 11:45 PM' post='140988']
can DNA heated upto 90C be used for electrophoresis?
if sonication or Rest. Enz. are not available, is there a way to fragment DNA and run them for viewing by gel staining?
i tried but couldnt get proper bands as i was told its because os denaturation of DNA.....
will heat treated dna wont give bands???
please help
[/quot
Hi there..
I am still learning thought, but as far as I am concern 90 degree at long hours will denature your DNA.
how big is ur fragment?
if sonication or Rest. Enz. are not available, is there a way to fragment DNA and run them for viewing by gel staining?
i tried but couldnt get proper bands as i was told its because os denaturation of DNA.....
will heat treated dna wont give bands???
please help
Depends on how long, generally not a big problem. DNA will renature upon cooling. It will give bands just fine.
Fragmenting :
Partial DNase treatment. Why would you want to do this, though?
Vigorous vortexing, if it is HMW DNA.
If you have chromatin, you can use Micrococcal nuclease.
if sonication or Rest. Enz. are not available, is there a way to fragment DNA and run them for viewing by gel staining?
i tried but couldnt get proper bands as i was told its because os denaturation of DNA.....
will heat treated dna wont give bands???
please help
Depends on how long, generally not a big problem. DNA will renature upon cooling. It will give bands just fine.
Fragmenting :
Partial DNase treatment. Why would you want to do this, though?
Vigorous vortexing, if it is HMW DNA.
If you have chromatin, you can use Micrococcal nuclease.
ok....so first i should heat the DNA...to fragment it...n then cool it for some time....then it will give bands? is that so?
will putting it on ice pack allow its renaturation?
if sonication or Rest. Enz. are not available, is there a way to fragment DNA and run them for viewing by gel staining?
i tried but couldnt get proper bands as i was told its because os denaturation of DNA.....
will heat treated dna wont give bands???
please help
Depends on how long, generally not a big problem. DNA will renature upon cooling. It will give bands just fine.
Fragmenting :
Partial DNase treatment. Why would you want to do this, though?
Vigorous vortexing, if it is HMW DNA.
If you have chromatin, you can use Micrococcal nuclease.
ok....so first i should heat the DNA...to fragment it...n then cool it for some time....then it will give bands? is that so?
will putting it on ice pack allow its renaturation?
I think they are is saying that heating will not fragment the DNA, only denature it (sstrand)
You might try vortexing it (as was suggested) or pipetting up and down vigorously to shear the DNA.
Hallo, you can fragment DNA by ultrasound and by exposition to UV 260nm, but by carefull to degradate it.
IN method SSCP single stranded conformational polymorphism DNA is denatureted by heat , chilled on ice immediatelly and run on gel
good luck