According to what I read, most of PCR applications are still possible if biotin is attached to the 5’- end of the oligonucleotide, but if the biotin is attached at the 3’- end, the modification usually block this end for further enzymatic reactions (nevertheless, some sensitive applications might detect elongation even of 3’-modified oligonucleotides).

Hoping I have helped you

Good luck

Thanks Marcello. According to the information of the Oligo-synthesizing company which I got today, the biotin is conjugated with the 5'-phosphate group at the 3'-end of the oligo. It should therefore work. I will see on Monday and let you know.

Best,

Jochen

I think Marecella is right. We offen used 5' biotinylated for LAMP and even for PCR. I don't think 3' biotinylated primers may not work for PCR amplification.

Hi,

Indeed, 3'-biotinylated oligos do not work in PCR as we just figured out. Thank you so much for your answers.

Best wishes,

Jochen