TY - JOUR
T1 - Phosphorothioated Primers Lead to Loop-Mediated Isothermal Amplification at Low Temperatures
AU - Cai, Sheng
AU - Jung, Cheulhee
AU - Bhadra, Sanchita
AU - Ellington, Andrew D.
N1 - Funding Information:
This work was supported by National Aeronautics and Space Administration Astrobiology Institute Cooperative Agreement Notice (NASA NAI CAN) [Grant NNX15AF46G] and Welch [Grant F-1654]. This publication was also made possible through the support of a grant from the John Templeton Foundation [Grant 54466]. The opinions expressed in this publication are those of the authors and do not necessarily reflect the views of the John Templeton Foundation. Funding for the open access was provided by the John Templeton Foundation.
Publisher Copyright:
© 2018 American Chemical Society.
PY - 2018/7/17
Y1 - 2018/7/17
N2 - Loop-mediated isothermal amplification (LAMP) is an extremely powerful tool for the detection of nucleic acids with high sensitivity and specificity. However, LAMP shows optimal performance at around 65 °C, which limits applications in point-of-care-testing (POCT). Here, we have developed a version of LAMP that uses phosphorothioated primers (PS-LAMP) to enable more efficient hairpin formation and extension at the termini of growing concatamers, and that therefore works at much lower temperatures. By including additional factors such as chaotropes (urea) and single-stranded DNA binding protein (SSB), the sensitivities and selectivities for amplicon detection with PS-LAMP at 40 °C were comparable with a regular LAMP reaction at 65 °C.
AB - Loop-mediated isothermal amplification (LAMP) is an extremely powerful tool for the detection of nucleic acids with high sensitivity and specificity. However, LAMP shows optimal performance at around 65 °C, which limits applications in point-of-care-testing (POCT). Here, we have developed a version of LAMP that uses phosphorothioated primers (PS-LAMP) to enable more efficient hairpin formation and extension at the termini of growing concatamers, and that therefore works at much lower temperatures. By including additional factors such as chaotropes (urea) and single-stranded DNA binding protein (SSB), the sensitivities and selectivities for amplicon detection with PS-LAMP at 40 °C were comparable with a regular LAMP reaction at 65 °C.
UR - http://www.scopus.com/inward/record.url?scp=85049643942&partnerID=8YFLogxK
U2 - 10.1021/acs.analchem.8b02062
DO - 10.1021/acs.analchem.8b02062
M3 - Article
C2 - 29968462
AN - SCOPUS:85049643942
SN - 0003-2700
VL - 90
SP - 8290
EP - 8294
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 14
ER -