Genomic Reagents FAQ: Trying to perform size selection with AMPure XP? Beckman Coulter Life Sciences has recommendations for you. Beckman Coulter Life Sciences is taking actions in the best interests of our associates, customers, and business partners as we navigate the growing threats of the 2019 Novel Coronavirus disease (COVID-19). AMPure XP is a flexible reagent suitable for manual or automated workflows. Below is estimated hands-on time and total time, in minutes, for AMPure XP. Scalable based on throughput; Quick transition with ready-to-implement methods; Knowledgeable support for reagents, automation and methods all from a single vendor. AMPure XP for PCR Purification Cleanup and Size Selection. You can use our proprietary SPRI paramagnetic bead-based chemistry to remove contaminants (dNTPs, salts, primers, primer dimers) throughout your NGS workflows. Used in a variety of NGS library prep chemistries; Compatible with manual and automated processing; High recovery of amplicons. AMPure XP 5 mL reagent kit is a highly efficient, easily automated PCR purification system that delivers superior quality DNA with no salt carryover. Requiring no centrifugation or filtration, AMPure XP can be easily used in manual and automated 96- or 384-well formats. NC9959336 $415.40 / Each.
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. 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.
Ampure Xp Beads Patent
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.
Beckman Ampure
Without any further ado, here are the results: