amplimer

amplimer

OLIGONUCLEOTIDE used as a PRIMER of DNA synthesis in the POLYMERASE CHAIN REACTION.
References in periodicals archive ?
Multiplex PCR amplimer conformation analysis for rapid detection of gyrA mutations in fluoroquinolone-resistant Mycobacterium tuberculosis clinical isolates.
A 535bp amplimer was generated with primers MTLa1-F (5'-TGAAAATGAAGACAATGCGA-3') and MTLa1-R (5'CATCTTTTTCTGCTATCAATTC-3') in the presence of MTL type a DNA, and a 615-bp product resulted from primers MTL[alpha]1-F (5'-ATGAATTCACATCTGGAGGC-3') and MTL[alpha]1-R (5'-CTGTTAATAGCAAAGCAGCC-3') in the presence of MTL type [alpha] DNA.
The second dystrophin amplimer group consists of 38 amplicons of symmetric exons [(0,0), (1,1), (2,2)].
The primer set Aca16Sf1010 (5'-TTATATTGACTTGTACAGGTGCT-3') and Aca16Sr1180 (5'-CATAATGATTTGACTTCTTCTCCT-3'), which was designed to give an amplimer of 161 bp, was used for detecting Acanthamoeba DNA (3).
We modified the molecular methods (7) by using different forward and reverse primers to lengthen the PCR product from a 99-bp to a 176-bp amplimer. We had difficulty resolving the 99-bp amplimer cut and uncut fragments from the 22 and 23mer PCR primers on 3% agarose, and the 176-bp product was more easily resolved, as was the 118-bp digestion product.
An important result was that, as expected, Inv22 Southern blot patterns (distal and proximal) yielded the same Inv22-specific amplimer (559 bp) without significant differences in relative signal intensities.
(The study was approved by the Institutional Ethical Committee, and informed consent was obtained from all participating individuals.) The PCR 206-bp product was amplified in 20 [micro]L containing 20 ng of genomic DNA, 0.5 [micro]M each amplimer, 100[micro]M deoxynucleotide triphosphates, 10 mM Tris-HCl (pH 8.3), 1.5 mM Mg[Cl.sub.2], 50 mM KCl, and 0.5 U of Taq polymerase (Platinum; Invitrogen) with the following conditions: initial denaturation at 94[degrees]C for 5 min; 40 cycles of denaturation at 94[degrees]C for 30 s, annealing at 59[degrees]C for 30 s, and extension at 72[degrees]C for 45 s; followed by a final extension at 72[degrees]C for 7 min.
This is achieved by covalently attaching one primer to the microwell before amplification, thereby allowing colorimetric detection of a solid-bound amplimer carrying a detector tag after washes.
Here we present the integrated system Three-STAR, the amalgam of three techniques using a STAR annealing function amplimer.
qRT-PCR reactions, and analysis of amplimer products were carried out accordingly to the methods already detailed [19].
All primer sequences were compared against each other and homology searches performed against the GeneBank database for sequence similarities using BLAST program (NCBI, USA).An amplimer of 232 bp for AHCYTOEN gene was obtained from both field and reference samples.
Finally, specificity of RT-PCR reaction was confirmed by determining the characteristic temperature of melting for each amplimer and by 6% polyacrylamide gel (PAA) electrophoresis of RT-PCR products with their visualization using silver staining.