I used DIG High Prime a few times and had no problems. However
it occurred to me that Random-Primed labelling leads to varying
sizes of probes leading to a potential for false positives. Hence I
switched to direct incorporation of DIG by PCR - much better,
quicker and easier! In my case it is also cheaper!
Mail me direct if you want any more info regarding this.
(No association with BM but if they want to fund my research........)
<color><param>0000,0000,FF00</param>neufeld at paprican.ca wrote:
>Hello, For quite some time I have been trying to label my two DNA gene probes
>(1.2KB, 1.9kb) with the Boehringer Mannheim product, DIG High Prime
>(random-primed labeling). However, I have been unable to label at an
>efficiency of more than 1/10 to 1/100 of the expected yield. I have also
>been trying to label the control unlabeled DNA that the company sends with no
>better success. More confusing is the variability of labeling efficiency
>from one labeling to the next. What I would like to know is if there is
>anyone at all who has ever used this product before? How did it work for
>you? If I cannot find anyone who has successfully used High Prime then I
>will probably attempt another DIG labeling method (any suggestions?). Thanks,
>Josh. P.S. Labeling success is measured by diluting out probe and spotting on
>a membrane with control-labeled DNA.
Raymond Chawla B.Sc.
University College Dublin
Belfied, Dublin 4, Ireland.
E-mail raymond.chawla at .ucd.ie
Phone (01) 706-1536