The NEXTflex[TM] Amplicon
Panels allow researchers to sequence up to 2,000 amplicons
using a simple workflow with 100% coverage of targeted regions.
Commercial kit to prepare DNA libraries from PCR amplicons
of HLA genes for the sequencing on the Illumina MiSeq next generation sequencing platform.
The subsequent high temperature allows the start of amplification by denaturing target strands for primer access (and in most cases, activating a "hot start" polymerase), and also acts to permanently inactivate the ung so it can't interfere with the production of new, uracil-containing amplicons
Approaches that can be used to circumvent this limitation are (a) amplicons
can be designed with specific [T.
TM)] High-Fidelity DNA Polymerase, that provides the highest fidelity amplification currently available, while also offering robust amplification across the broadest range of amplicons
, including those that are GC- and AT-rich.
Primers should be redesigned for amplicons
that do not amplify sufficiently well despite optimization.
The authors amplified genomic material from swine sera using primers based on hepatitis E virus (HEV) genomic sequences, and sequenced the amplicons
The sample preparation, DNA extraction, PCR setup, electrophoretic separation, and sequencing of amplicons
are described elsewhere (6,9); however, we used 0.
Clear and intense amplicons
(bands) in the gel were considered to reflect that the DNA used was of high quality and quantity.
Amplification of the product specific for BYVaV was confirmed by cloning and sequencing selected amplicons
This 74-bp sequence is completely homologous with the alpha-2 microglobulin accession number X14552 and J00738 sequences, qRT-PCR of an amplicon
overlapping this 74-bp sequence, as well as for the alpha-2 microglobulin sequences X14552 and J00738, showed a substantial reduction (> 100-fold) in message levels for the day 7 high-dose methapyrilene sample, whereas a similar analysis of amplicons
designed for the amplification of U18419 and AL180288 showed induction levels above 5-fold (Figure 2).
As part of the agreement, Avalon will provide Aventis with a subset of its library of more than 200 identified amplicons
(regions of chromosomal amplification) that have been discovered through cytogenetic analysis.