Capture and Elute Step for Control DNA (E2600)

Introduction

Included in the kit are control DNA and primers to detect a methylated region, LINE element, an active gene exon, RPL30, and a micro RNA control region, MirA from a sample of fractionated HeLa DNA. (Sequences of the primers and target DNA are listed in the appendix section). A typical control reaction, that will yield a sufficient quantity of eluted material to perform qPCR reactions in triplicate, is outlined below.

Protocol

1. Perform capture experiment using 5 µl (0.5 µg) of fractionated HeLa Control DNA and 15 µl MBD2-Fc/Protein A Magnetic Beads as described above.
2. Elute the beads by incubation at 65°C in 100 µl for 15 minutes of DNasefree water