AMPURE® PURIFICATION SYSTEM
AMPure is a high recovery, SPRI™-based PCR clean up system capable
of purifying amplicons as small as 100bp. AMPure yields high quality PCR
amplicons using a simple, three-step protocol requiring no centrifugation
or filtration steps. The protocol can be easily automated or performed
manually in both 96 and 384-well formats.
EFFICIENT PCR PURIFICATION
Efficient removal of unincorporated dNTPs, primers and salts
High recovery of small and large amplicons
High quality PCR amplicons
AMPure is flexible enough to be performed in a variety of formats for an
array of applications:
Rolling Circle Amplification
EFFECTIVE CLEAN UP
AMPure removes unincorporated primers, dNTPs and salts used during
PCR amplification. These products can interfere with downstream applications.
AMPure routinely has recovery rates greater than 90% for amplicons
larger than 200bp and greater than 85% for smaller PCR amplicons,
such as the 165bp fragment shown in Figure 2.
HIGH PURITY – HIGH PERFORMANCE
AMPure-purified samples are ideal sequencing templates. The table
below shows sequencing results from a single 976bp PCR template purified
using AMPure or ExoSAP-IT™. The average Phred20 read length and average
pass rate for the AMPure-purified samples were 701 and 100% respectively.
ExoSAP-IT, an alternative approach of inactivating polymerases and
degrading unincorporated primers following PCR thermo-cycling, produces
samples with lower purity due to the presence of salts and degraded
enzymes and primers. Sequence results following ExoSAP-IT digestions
are consistently inferior, with fewer Phred20 bases per read and lower
pass rates. Figure 3 exhibits the superior sequencing performance
attained using AMPure.
AMPure produces reliable recoveries of both large and small amplicons.
Figure 4 shows two gels following AMPure purification of 967 bp and
165 bp PCR fragments.
AMPure demonstrates excellent recovery rates of both small and
large amplicons while maintaining high sample purity. AMPure samples
have little to no contamination as shown by the lack of smearing above
the bands (Figure 5). In contrast, alternative purification products
have decreased recovery rates and/or lower sample purity, which affect
the performance of downstream applications such as sequencing.
COMPLETE AMPURE AUTOMATION
The lack of centrifugation and filtration steps allows AMPure protocols
to be completely automated while maintaining superior performance. AMPure
is fully compatible with most commonly used low, medium and high throughout
liquid handling systems. Contact Agencourt for the most up-to-date list
of supported platforms.
AMPure has been successfully automated on the Biomek FX from Beckman Coulter
in 96 and 384-well formats. (Photo courtesy of Beckman Coulter.)