Protocol Online logo
Top : New Forum Archives (2009-): : Molecular Biology

Can you see any digestion? - (Jul/25/2010 )

Hello,
Would any one please tell on which samples the digestion is occuring? the products are 107 and 82bp..
Attached File

Attached File

-Fhannan-

Its the same gel, but just 2 pictures?

And you see a band near 100bp in every lane... but is this digested?

Where is your negative control? (the sample without digestion?)

-pito-

No these are different samples..
I dont load a -ve control as the presence of only the 107 bp without the 82bp should work as -ve control...
i presume the last four wells of pic 1 are not showing any digestion and should be considered as the -ve control..
my suspects are the shadow bands, do they represent a +ve reaction where digestion is taking place???

-Fhannan-

Fhannan on Sun Jul 25 15:43:28 2010 said:


No these are different samples..
I dont load a -ve control as the presence of only the 107 bp without the 82bp should work as -ve control...
i presume the last four wells of pic 1 are not showing any digestion and should be considered as the -ve control..
my suspects are the shadow bands, do they represent a +ve reaction where digestion is taking place???


If you did load a negative control... you would not have this problem.
And the be honest: I see no difference in pic 1 between the last 4 and first 4 lanes.. But this can be due the low quality of the pic I am seeing here when I zoom in.


and what is a positive controle?? You allready have a sample that has been digested before???
In the +ve lane, I hardly see anything at the +-100bp ?

-pito-

Sorry separation on this gel isn't good enough. What kind of gel are you using?

-perneseblue-

12% PAGE, the lower bands are primer dimmer...my question is should the digested bands appear as intense as the undigested band on the gel? or i can consider the faint band as the lower digested product?

-Fhannan-