Enhanced solid phase PCR: mechanisms to increase priming by solid support primers

Author(s): Khan, Z; Poetter, K; Park, DJ;
Details: Publication Year 2008-04-15,Volume 375,Issue #2,Page 391-393
Journal Title: ANALYTICAL BIOCHEMISTRY
Publication Type: Journal Article
Abstract: Conventional solid phase amplification regimens such as solid phase PCR (SP-PCR), asymmetric SP-PCR, and bridge PCR are mechanistically limited with respect to amplification efficiency and solid support primer involvement. Here we present enhanced solid. phase PCR (ESP-PCR) in which solid support primer priming is facilitated by its nesting and high melting temperature (T.) relative to the aqueous counterpart. In the study, we demonstrated increased solid support surface loading using ESP-PCR versus standard SP-PCR for three diagnostic targets: Neisseria gonorrhoeae opa (9.89-fold), N. gonorrhoeae pilS (2.14-fold), and Chlamydia trachoinatis cryptic plasmid orf3 (1.41-fold). Furthermore, we applied ESP-PCR to detect five copies of N. gonorrhoeae and C trachomatis DNA. (c) 2008 Elsevier Inc. All rights reserved.
Publisher: ACADEMIC PRESS INC ELSEVIER SCIENCE
Keywords: POLYMERASE-CHAIN-REACTION; ASYMMETRIC PCR; AMPLIFICATION; HYBRIDIZATION; GENOME; GENE
Publisher's Version: https://doi.org/10.1016/j.ab.2008.01.021
Terms of Use/Rights Notice: Refer to copyright notice on published article.

Creation Date: 2008-04-15 12:00:00