REPLI-g Mini Kit (25)

Designed for highly uniform whole genome amplification, QIAGEN's REPLI-g Mini Kit addresses limitations imposed by insufficient quantities of genomic DNA. The kit uses Multiple Displacement Amplification (MDA) technology with Phi29 polymerase in an isothermal reaction. According to the manufacturer, Phi29 polymerase can replicate up to 70 kb without dissociating from the genomic DNA template, has 3'→5' exonuclease proofreading activity and maintains up to 1000-fold higher fidelity than Taq DNA polymerase.

The kit accepts genomic DNA, blood, cells or tissue as starting material, with input amounts of >10 ng purified genomic DNA, or 0.1–0.5 µl whole blood (>600 cells/µl). DNA yields consistently reach 10 µg per 50 µl reaction regardless of template quantity. Reaction time is 10–16 hours, hands-on time is 15 minutes, and the protocol uses an alkaline denaturation step in a tube format with a minimal pipetting volume of 0.5 µl.

The amplified DNA is suitable for downstream applications including genotyping, hybridization, RFLP, end-point PCR, quantitative real-time PCR, microarray and NGS. QIAGEN states that average product length is typically greater than 10 kb, with a range between 2 kb and 100 kb. The amplification is highly uniform across the entire genome with minimal sequence bias, and amplified DNA can be stored at –20°C without negative effects.