Miniprimer PCR (CAT#: STEM-MB-0213-WXH)

Characterization of 16S rRNA gene sequences has become a central feature of microbial ecology. as the 16S rRNA sequence database has grown, it has become evident that many sequences deviate within the most conserved regions targeted by “universal” 16S rRNA gene PCR primers. To accommodate these deviations, commonly used primers have been modified with degenerate positions to enable the primers to target a wider range of 16S rRNA gene sequences. However, because polymerases used for PCR require primers of ∼20 to 30 nucleotides(nt), 16S rRNA gene primer design has been constrained to target conserved regions of those lengths. New thermostable polymerases have recently become available, opening the possibility of changes in primer design. Preliminary data suggested that some of these polymerases may be able to utilize primers shorter than the minimum length of ∼20 to 30 nt typically recognized by the standard enzyme used for PCR, Taq DNA polymerase.

Broaden the detectable scope of 16S rRNA sequences, which benefits the study of microbial ecology.

1.DNA extraction
2.Primer design
3.Miniprimer PCR amplification
4.Data analysis

10-nucleotide “miniprimers”