Taking into consideration that my DNA can be prepared in low quantities (~35ng/micro L), average final volume 50 micro L. I have tried many protocols as almost them require much higher quantities (milligrams). I would request your help. I am asking for a detailed procedure, please.
You could order one primer labelled with biotin at its 5'end and use about 23 bases at the other end of the dna fragment and then pcr and the pcr product will be biotin labelled.
I'd give Terminal Transferase a try. It adds dNTPs to the 3' end of DNA and works with modified dNTPs and works with ng amounts of DNA. You can play around with the dNTP concentration to change the number of them added to your DNA ends.
Check NEB for a detailed protocol
To conjugate dsDNA (150bp) with biotin, you need to use a biotinylation kit. The kit typically includes a biotinylation reagent, a buffer, and a cleavable linker. After the dsDNA is mixed with the biotinylation reagent, it is incubated at room temperature for a period of time. The biotinylated dsDNA is then separated from the unreacted reagent by gel electrophoresis, and the biotinylated dsDNA is recovered from the gel.
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