Direct Oral Swab 2xPCR Master Mix

Direct Oral Swab 2xPCR Master Mix

  • DNA isolation is not needed anymore -
    Direct Oral Swab 2x PCR Master Mix allows you direct PCR without the tedious steps of any previous DNA extraction steps. Simply run your genotyping or pathogen detection assays directly from saliva or buccal swab samples.

    Direct Oral Swab 2x PCR Master Mix contains HiDi Taq DNA polymerase and thus, is also proficient in all assays in which high single nucleotide discrimination is required, for instance in allele-specific amplifications (ASA) by PCR. HiDi Taq DNA polymerase efficiently amplifies from primers that are matched at the 3'-end and discriminates primers that are mismatched.

    The kit comes together with an optimized buffer system and a separate lysis buffer for saliva samples.

  • - Direct PCR from saliva samples
    - Pathogen detection directly from saliva samples
    - Genotyping & genomic profiling
    - SNP-detection by allele-specific amplification (ASA) / Allele-specific PCR - directly from saliva samples
    - HLA genotyping
    - End-point PCR
    - Real-time PCR with with fluorescence-based hydrolysis probes

    All HiDi products are also compatible for real-time cycling, when adding a suitable real-time dye, for example GreenDye (#2000) or a fluorescent probe.

    Direct Oral Swab 2xPCR Master Mix can also be used for hydrolysis-based probes (e.g. TaqMan) as the 5'-3' nuclease.

    We recommend designing primers with a short amplicon length (about 60-200 bp) for best results, but also longer amplicon lengths are possible. The addition of additional magnesium chloride (+ 0.5 - 1.5 mM) might be needed in case of longer amplicons >500 bp.

  • PCR activity: Direct Oral Swab 2x PCR Master Mix was tested for successful allele-specific PCR performance detecting a genomic SNP (rs72921001) in human genomic DNA from saliva samples. PCR products were subsequently analysed on a 2.5% agarose gel. A specific product is visualized by ethidium bromide staining at the right amplicon length of 109 bp for the matched primer. In the case of the mismatch primer no product formation is visible after 40 cycles.

    DNA polymerase activity: HiDiTaq DNA polymerase activity has been monitored and adjusted to a specific DNA polymerase activity using an artificial DNA template and a DNA primer.

    Enzyme-concentration has been determined by protein-specific staining. Please inquire more information at info@mypols.de for the lot-specific concentration.

    No contamination has been detected in standard test reactions.

Example Primer Design

Matching vs. mismatching nucleotide is placed at the 3'-end of the primer for best discrimination results.

Example Results - There´s no accounting for taste

Cilantro: some people love it in their food, some hate it. Here we are detecting a genomic SNP (rs72921001) in HeLa genomic DNA. This SNP is reported to be close to a number of genes coding for olfactory receptors. (Reference: Eriksson N. et al. (2012), "A genetic variant near olfactory receptor genes influences cilantro preference.")

Considering, that only the C-allele specific primer is extended and yielding in a specific amplicon, we can conclude a genetic predisposition in disliking cilantro, as this SNP is significantly associated with detecting a soapy taste to cilantro.  

Allele-specific PCRs were performed from 1 ng/µl of HeLa gDNA in the presence of a realtime dye, indicating the amplification of the C-allele specific primer only. The A-allele specific primer is discriminated, thus not amplified up to 50 cycles.

PCR products were subsequently analysed on a 2.5% agarose gel. Specific product is visualized by ethidium bromide staining at the amplicon length of 109 bp.


Related Items