Hi,
> > I'm planning to use slot blot hybridization to calculate the gene
> > copy number in the genome. However, my slot blot give a very weak signal
> > even 50ug of genomic DNA was applied. Is there any possible casue for
> > this..?? I am using DIG labelling and chemiluminecence detection. The
> > system works fine for southern blot (using the same probe and genomic
> > DNA), which can detect a single copy gene using 5 - 10ug of genomic DNA.
> > So, I don't know what's wrong with the slot blot. Can anyone give any
> > suggestion??
>> Are you denaturing your genomic DNA before you apply it to the filter?
>
Yes, by adding NaOH to final conc. of 0.4N and boil for 16 minutes.
However, after the slot blot process, I notice something stick on the
membrane. This is only observed in slot with high amount of DNA (e.g. 35ug) Is
that the DNA..?? And how should I fix the DNA on the membrane??
Regards,
Lum
