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.

Sample Preparation

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

Pricing is sample number dependent. Please contact Laura Kasch for pricing.

Ordering

Please place your order through our secure online ordering system. See FAF Order Forms: FAF Whole Genome Amplification.