: : FP-TDI Technology

: : SNP Education

: : External FP-TDI Resources

FP-TDI Technology

Contents:
Assay Description
High-throughput capabilities
Scoring FP-TDI data
Resources for additional information

Our laboratory uses a genotyping method for SNPs that combines the specificity of nucleotide incorporation by DNA polymerase and the sensitivity of fluorescence polarization, known as Template-directed Dye Terminator Incorporation assay with Fluorescent Polarization detection (FP-TDI). Following PCR amplification of the target region, a SNP-specific primer anneals immediately upstream of the polymorphic site in the target DNA. Appropriate dye-labeled terminators extend the SNP-specific primer by one base; by determining which terminator is incorporated, the allele present in the target DNA can be inferred.

The highly specific and sensitive nature of FP-TDI allows us to run assays in a 384-well plate format, generating about 10,000 genotypes per day. During our participation in the International HapMap Project , we developed a Laboratory Information Management System (LIMS) and perfected experimental protocols to support high throughput SNP genotyping.

Already use FP-TDI? Patricia Taillon-Miller from our lab put together a guide to Scoring your FP-TDI Data.

PerkinElmer offers excellent informational pages on SNPs and FP-TDI.

These publications were the first to describe the FP-TDI technique:

[1] Chen, X., Levine, L. and Kwok, P-Y., “Fluorescence polarization in homogeneous nucleic acid analysis”, Genome Res. (1999) 9,492-498

[2] Hsu, T.M., Chen, X., Duan, S., Miller, R.D., and Kwok, P-Y., “Universal SNP Genotyping assay with fluorescence polarization detection”, Biotechniques (2001) 31:560-570.