Use of the DCode System to Detect the Food-Borne Bacterial Pathogen Listeria monocytogenes
... gene of L. monocytogenes, Listeria
innocua, and Listeria seeligeri by the polymerase chain reaction (PCR)*
using primers that anneal
to conserved sequences, yielded multiple copies
of gene fragments that, when evaluated by density gradient gel electrophoresis
Rapid, Reproducible Real-Time Quantitative RT-PCR Using the iCycler iQ Real-Time PCR Detection System and iQ Supermix, Rev A
... II reverse transcriptase (Invitrogen) according to the
Gene-specific primers (GSPs) were designed to anneal
near the 3' end
of two mRNA transcripts and to generate PCR products 75300 base pairs
long. Three GSP pairs amplify different ...
RNA Amplification for Array Analysis
... the typical probe orientation
for microarrays? Oligonucleotides are single-stranded and
therefore only anneal
to their complement. Probes for
microarrays are either generated from an RT reaction (cDNA) or
from linear ...
Thermal Cycling Profile for Standard PCR
heating of the PCR mixture for 2 minutes at 9495C is enough to completely
denature complex genomic DNA so that the primers can anneal
to the template
as the reaction mix is cooled. If the template DNA is only partially denatured,
it will tend to snap-back very ...
... the required primers (see figure below).
Make primers that anneal
to sequences in exons on both sides of an intron
(Panel 1). With these primers, any product amplified from genomic DNA
will be ...
QIAGEN Multiplex PCR Handbook
... of the primer mix.
Primers may anneal
to pseudogene sequences and additional
PCR products may be amplified. Review primer design to avoid detection
Using the TripleMaster PCR System for Robust Amplification of
GC-Rich DNA Templates
... renaturation of the template
and the formation of secondary structures within each strand before the
primers have a chance to anneal
to their specific template sites. These
properties of GC-rich DNA sequences lead to inefficient primer-template
annealing and ...
Expression and Purification of Recombinant Proteins That Have Native Amino
... that are complementary
to the single-stranded tails of the pCAL-n-EK vector. The vector and insert
DNA are combined, allowed to anneal
at room temperature and transformed
into highly competent bacterial host cells. The resultant colonies can
then be screened for ...
High-Level Protein Expression, One-Column Purification, and FLAG
Epitope Tagging in E. coli
... is purified, then
treated with Pfu DNA polymerase in the presence of dATP. The vector
and insert DNA are combined, allowed to anneal
at room temperature, and
transformed into highly competent bacterial host cells. The resultant
colonies can then be screened ...
Map and Link Human Genetic Disorders with SSLP Analysis
... location. 1 Often these
markers are (CA)n repeat polymorphisms. 1 These polymorphisms can
easily be genotyped by PCR with primers that anneal
to single-copy DNA, which
flanks the repetitive element.
Genotyping, using microsatellite markers, at specific locations throughout
Performance Comparisons of Commercial RT-PCR Systems
... and RNA (200
ng of total RNA). The reaction mixtures were incubated at 65C for 5 minutes
and cooled to room temperature to allow the primers to anneal
to the RNA.
MMLV-RT (10 U) was added, and cDNA synthesis was carried out at 37C for 15
minutes (0.3- and 1.8-kb targets) or 30 minutes (7.6-kb ...
prostar RT-PCR Systems for Robust High-Fidelity RNA Amplification
... ng of bacteriophage MS2 RNA). The reaction mixtures
were incubated at 65C for 5 minutes, then cooled to room temperature to allow
the primers to anneal
to the RNA. A total of 10 U of MMLV-RT was added, and cDNA
synthesis was carried out at 37C for 15 minutes (targets <2 kb) or 30
minutes (targets ...
Pfu DNA Polymerase as a Hot Start Alternative
... polymerase at lower temperatures
( figure 2 ), Taq DNA polymerase has a greater potential for extending
mispaired primers that anneal
during these three low-temperature phases
of PCR amplification.
Hot start PCR protocols have been developed to reduce or prevent ...