The SNaPshot Multiplex System is a primer extension-based method developed by Applied Biosystems for multiplexed genotyping of up to 12 SNPs. A simplified workflow is shown as below: Multiplex PCR is used to obtain genomic regions containing SNPs of interest. Next, unlabelled oligonucleotide primers with different lengths representing different SNP loci as well as fluorescently labelled ddNTP are mixed with template DNA (multiplex PCR products). Each of these oligonucleotide primers contains sequence complementary to one of template DNAs and stops one nucleotide before the SNP. The polymerase extends the primer by one nucleotide, incorporating a single ddNTP to its 3 ́ end which is complementary to the SNP. The fluorescence color readout indicates which base was added.
Technique Highlights
– High accuracy SNaPshot also called minisequencing, the accuracy of which matches sanger sequencing.
– Multiplexing capability SNaPshot simultaneously interrogates up to 12 SNPs in a single reaction as apposed to only
one SNP per reaction with Taqman or traditional PCR-based approaches.
– Sensitive allele-frequency detection SNaPshot can detect rare allelic genotypes comprises as low as 5% of the
total DNA while sanger sequencing can only detect alleles when they comprise >20% of a sample.
Sample Requirement
Sample Type: Genomic DNA
Amount: DNA >= 1 ug; concentration >= 20 ng/ul
Purity: OD260/280 = 1.7~2.0
Sample without severe degradation
Service Workflow
1. Cusomtized sequencing experimental design
2. Sample receiving
3. DNA extraction and QC
4. SNaPshot multiplex genotyping
5. Project final report
Deliverables
1. SNP genotyping analysis result
2. QC result
