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Anneal in Biological Technology

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 manufacturers instructions. 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 ...

Primer design

... 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. 9. Amplification of pseudogene sequences Primers may anneal to pseudogene sequences and additional PCR products may be amplified. Review primer design to avoid detection of ...

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 Acid Sequence

... 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 the ...

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 ...
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