Whole genome amplification (WGA) of extracted DNA is carried out using a REPLI-g kit (Qiagen). The kit uses Multiple Displacement Amplification technology to produce highly uniform amplification across the genome. The genome is isothermally amplified utilizing a DNA polymerase with 3′ – 5′ exonuclease proofreading activity capable of replicating up to 100 kb without disassociating from the DNA template. Typical yields from the REPLI-g kit are 40ug DNA with a product range of 2 to 100 kb and an average of greater than10 kb. Poor quality genomic dna may result in synthesis failure, reduced call rates in downstream applications, or loss of loci or alleles in the amplification product.
We accept genomic DNA, whole blood, frozen blood, packed cells (buffy coats), blood spots, and cultured cells for whole genome amplification. Samples are amplified using Qiagen’s REPLI-g Midi Kit. OD is determined by pico green analysis using a Spectra Max Gemini XS fluorometer. Concentrations will be returned to you in an Excel document. See below table for specifics on amount to send for each sample type and shipping conditions.
Sample Type Amount Required Shipping Conditions
Genomic DNA 100ng at 20-100 ng/µl in TE or water Ice packs (tubes), dry ice (plates)
Whole Blood* 5 to 50 µl Ice packs
Frozen Blood* 5 to 50 µl Dry ice
Packed Cells* (buffy coat) 5 to 50 µl Dry ice
Blood Spots Two punches, 2-3mm diameter RT or dry ice if currently stored in a freezer
Cultured Cells >600 cells in PBS Dry ice
* Whole and frozen blood, and packed cells should be preserved with EDTA or Sodium Citrate.
Pricing is sample number dependent. Please contact Laura Kasch for pricing.
Please place your order through our secure online ordering system. See FAF Order Forms: FAF Whole Genome Amplification.