Can anybody suggest working primers for CONVENTIONAL PCR (not a real-time PCR) to detect Fusobacterium necrophorum and Bacteroides melaninogenicus? Data on PCR conditions and product size (in bp) are also necessary.
Thank you in advance!
Dear Modestas,
for Fusobacterium necrophorum you can use this:
Specific detection and differentiation of two subspecies of Fusobacterium necrophorum by PCR (Narongwanichgarn et al. 2003)
and for Bacteroides melaninogenicus you can try the setup described here:
Detection of Bacteroides fragilis in Clinical Specimens by PCR (Yamashita et al. 1993)
As a third option you can use 16S rRNA universal primers to generate a PCR product suitable for sequencing:
GM3 up: 5’-AGAGTTTGATCMTGGC-3’ and
GM4 lo: 5’-TACCTTGTTACGACTT-3’ (Muyzer et al. 1995)
Amplifying regards
Thomas
Dear Modestas,
for Fusobacterium necrophorum you can use this:
Specific detection and differentiation of two subspecies of Fusobacterium necrophorum by PCR (Narongwanichgarn et al. 2003)
and for Bacteroides melaninogenicus you can try the setup described here:
Detection of Bacteroides fragilis in Clinical Specimens by PCR (Yamashita et al. 1993)
As a third option you can use 16S rRNA universal primers to generate a PCR product suitable for sequencing:
GM3 up: 5’-AGAGTTTGATCMTGGC-3’ and
GM4 lo: 5’-TACCTTGTTACGACTT-3’ (Muyzer et al. 1995)
Amplifying regards
Thomas
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