How to measure biotinylated DNA? - (Jun/12/2006 )
i label my dna fragment with biotin
i use taq pol to elongate over biotin
after amplification, how i measure biotinylated dna? i know that biotin interfere with BET
can i measure the biotinylated dan on spectro?
thanks in advance
I don't understand? why did you not label your DNA after biotinylation? or, does your amplification scheme incorporate biotinylation?
if I am getting you clearly, you have product in the tube...the original template is biotinylated and the new product is not, and you want to know how much biotinylated is left? is this right?
try to PCR them if you wanna have purified DNA without trouble
excuse me but my english is very bad and i have trouble to explain me in this language
my original product is not label with biotin, i incorporate biotin in my dna fragment during the amplification (dUTP-biotin)
before to use this labeled dna fragment in my macro-array system, i would to quantify it
and i don't know how i do it
What you really care about is whether you can detect the biotin in your probe. Spot your probe on a membrane, and run your detection method on the membrane to determine the lower limit of detectability of the probe/detection technique.
In general, I've found that DIG is easier to work with than biotin labels, but that doesn't mean you will have any problems.