Available, the Agencourt AMPure XP reagent system is a highly efficient, easily automated PCR purification system that delivers superior quality DNA with no salt or dNTP carryover. The AMPure XP reagent system uses an optimized buffer to selectively and reversibly bind PCR amplicons that are 100bp and larger to solid-phase paramagnetic beads. AMPure XP clean-ups. AMPure XP can be performed either manually or automated on a liquid handling system. Difference in time between manual and automation is indicated. NR = Not Recommended For use in manual or automated methods based on batch size or overall throughput. The Agencourt AMPure XP PCR purification kit can be used in 96 and 384 well format. The following table illustrates the number of PCR reactions an Agencourt AMPure XP kit will purify depending on the format required by the user. AMPure XP AMPure XP Product Product # AMPure XP 5.0mL A63880 AMPure XP 60 mL A63881 AMPure XP 450 mL A63882.
I wanted to talk a little about the selection characteristics of Agencourt’s AMPure beads, a bead-reagent combination that purifies PCR reactions.
This stuff is incredible in terms of simplicity, efficiency, and high-throughput compatibility. I have a sneaking suspicion that AMPure, not unlike fire to Prometheus, was handed down from the gods to benefit humanity. Disfear torrent discography garth. You just dunk it into your sample, slosh it around, stick it to a magnet, wash, wash again, and elute in your favorite buffer. No muss, no fuss.
We were wondering, though, about its selection process. What size fragments are selected by the AMPure beads, specifically at which ratio of beads to sample? So, like diligent scientists, we rolled up the sleeves of our labcoats and… read the protocol.
The protocol recommends washing your sample in a 1.8:1 ratio of beads to sample, although it says that fragments less than 100bp will be omitted at this ratio, it doesn’t say which sized fragments will be selected. We found this remarkably helpful technical bulletin, which describes calibrating each batch of AMPure beads with various ratios of DNA ladder.
So I did our very own calibration with AMPure beads using Fermentas’s GeneRuler™ Low Range DNA Ladder (25-700 bp). I added 30ul ladder to various concentrations of AMPure beads according to Agencourt’s instructions.
(Actually, if you’re looking for good AMPure instructions, I recommend looking at Illumina’s TruSeq™ Sample Preparation Guide. Honestly, their instructions are more comprehensive than Agencourt’s, and easier to read.) After purifying each sample, I bookended the various AMPure:ladder ratios with 10ul non-purified ladder on a 2% TBE gel for easy comparison.
Ampure Xp Manual
Without any further ado, here are the results: