EBspot - (Sep/08/2006 )
I am very new to using EB spot to roughly quantitate DNA. I want to compare the intensity of my spots in the second and third rows with the first row to check whether my DNA concentration is correct. I have a feeling that my 150ug/ul spot is not correct and looks less bright than the 100ug/ul. What do you think?
Can anyone share with me the tips to estimate concentration from EBspot (the only method that I can use in my lab because we don't have nanodrop)? I find it very hard to determine the concentration from the intensity of the spots.
Thank you.
I am not an expert in EBspot. actually I dont know what it is.
But u say u r using it to determine DNA conc. , might as well.
Well, we just compare dif. DNA bands (on a gel) and judge by the intensity of bands. It migth not b very accurate but to have a rough estimate, its useful. I still find it hard, but with practice u will b fine.
Sample 1 : seems like 10ng
sample 2: approx. 50ng (based on top row), may b slightly less than 50.
But u say u r using it to determine DNA conc. , might as well.
Well, we just compare dif. DNA bands (on a gel) and judge by the intensity of bands. It migth not b very accurate but to have a rough estimate, its useful. I still find it hard, but with practice u will b fine.
Sample 1 : seems like 10ng
sample 2: approx. 50ng (based on top row), may b slightly less than 50.
Comparing the top row and second row of dots, do you think the dots in the second row are of the right concentration as stated? Thank you.