I wanted to talk a little about the selection characteristics of Agencourt's AMPure beads, a bead-reagent combination that purifies PCR reactions.
Scanwizard pro 7 mac download. 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.
About This Manual The information in this manual is organized as follows: Agencourt AMPure XP PCR Purification Provides an overview of the Agencourt AMPure XP PCR. purification process, materials such as consumables and reagents needed, and the procedure for either a. ICELL8 Human TCR a/b Profiling User Manual (072219) takarabio.com Takara Bio USA, Inc. Hay day mac download. Page 7 of 61 - Figure 2. Protocols for ICELL8 Human TCR a/b Profiling. Library Validation and Quantification (Section IV.L) Purification of Amplified Libraries Using Agencourt AMPure XP Beads (Section IV.K). Marsh AB-0937), but will require manual intervention to move the plates on and off the Agencourt SPRIPlate384. Gently shake the Agencourt AMPure bottle to resuspend any magnetic particles that may have settled. Add Agencourt AMPure according to the following PCR reaction volume chart: PCR Reaction Volume (µL) Agencourt AMPure Volume (µL) 5 9.
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. Download pixlr mac.
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 Beads Beckman Protocol
Without any further ado, here are the results:
