In addition to Real Time PCR methods, Arizona Genetics Core offers SNP genotyping on the Agena Bioscience MassARRAY® MALDI-TOF instrument. Using this technology we can multiplex SNPs for up to 36 loci.
General Information
Sample Input
Each reaction requires 1µL of 10ng/µL DNA.
The DNA can be submitted in 96-well or 384-well plates with at least 2 open wells for negative controls
Expected Results
Genotypes are determined through ratio analysis of the MALDI-TOF mass spectrometer.
Details
Sample Preparation Details
- The DNA samples should be eluted in RNase free or HPLC water. The instrument is very sensitive to salt, so please avoid TE buffer.
- Arizona Genetics Core can extract and prepare your samples for SNP genotyping.
- The instrument utilizes 384-well plate format.
- The DNA can be submitted in 96-well or 384-well plates with at least 2 open wells per every 384 for negative controls.
- Each reaction requires 1µL of 10ng/µL DNA. It is best if 260/280 ratio is between 1.7 and 2.0.
Workflow
Contact Arizona Genetics Core to discuss SNPs of interest and multiplex design
Multiplex Design Information: This instrument can perform up to a 36-plex, but we typically achieve between 25 and 30 loci. To optimize the largest multiplex possible, it is best to provide 40-50 loci with specific notes about their relative importance. Provide the SNP ID (rs#) and the sequence. Within the sequence the SNP should be denoted in brackets (i.e. [G/A]), with 100-200 bases of flanking sequence on either side. Tri-allelic SNPs are also detectable. Multiplex PCR assays target ~100bp amplicons containing SNPs of interest.
If there are other known polymorphisms within the sequence please denote them as 'X' so we can design the primers around them. There cannot be polymorphisms bracketing the SNP of interest within 20bp; however, if there is a polymorphism upstream of the SNP of interest, we can design the primer on the antisense strand.
Three primers are utilized per SNP, two for amplification and one for the extension reaction. Primers are unlabeled and usually 15-25 bp in length.
Price
Academic (UA): $1400.00| Academic (non-UA): $1680.00 | Industry: $2100.00 Unit: 96-well Plate
Academic (UA): $2750.00 | Academic (non-UA): $3300.00 | Industry: $4125.00 Unit: 384-well Plate
Turnaround Time
Varies
Related Services
DNA Extraction
Real Time PCR (alternative SNP genotyping platform)
Additional Information
The price listed above is per 96-well or 384-well plate. If multiplexed at 30 SNPs cost ~ $0.20 per SNP
Primers are not included in the above pricing. These typically run ~$14/SNP.
For more information about the Agena Bioscience MassARRAY® system, please visit our FAQ page or go to http://agenabio.com/