Helicase‐Dependent Isothermal Amplification of DNA and RNA by Using Self‐Avoiding Molecular Recognition Systems
Identifieur interne : 001984 ( Main/Exploration ); précédent : 001983; suivant : 001985Helicase‐Dependent Isothermal Amplification of DNA and RNA by Using Self‐Avoiding Molecular Recognition Systems
Auteurs : Zunyi Yang [États-Unis] ; Chris Mclendon [États-Unis] ; Daniel Hutter [États-Unis] ; Kevin M. Bradley [États-Unis] ; Shuichi Hoshika [États-Unis] ; Carole B. Frye [États-Unis] ; Steven A. Benner [États-Unis]Source :
- ChemBioChem [ 1439-4227 ] ; 2015-06-15.
Abstract
Assays that detect DNA or RNA (xNA) are highly sensitive, as small amounts of xNA can be amplified by PCR. Unfortunately, PCR is inconvenient in low‐resource environments, and requires equipment and power that might not be available in these environments. Isothermal procedures, which avoid thermal cycling, are often confounded by primer dimers, off‐target priming, and other artifacts. Here, we show how a “self avoiding molecular recognition system” (SAMRS) eliminates these artifacts and gives clean amplicons in a helicase‐dependent isothermal amplification (SAMRS‐HDA). We also show that incorporating SAMRS into the 3′‐ends of primers facilitates the design and screening of primers for HDA assays. Finally, we show that SAMRS‐HDA can be twofold multiplexed, difficult to achieve with HDA using standard primers. Thus, SAMRS‐HDA is a more versatile approach than standard HDA, with a broader applicability for xNA‐targeted diagnostics and research.
Primers containing self‐avoiding molecular recognition system (SAMRS) components can eliminate artifacts in helicase‐dependent isothermal amplification (HDA)—artifacts that are common in classical HDA that uses standard primers. SAMRS therefore greatly expands the versatility of HDA, and supports low levels of multiplexing.
Url:
DOI: 10.1002/cbic.201500135
Affiliations:
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Unfortunately, PCR is inconvenient in low‐resource environments, and requires equipment and power that might not be available in these environments. Isothermal procedures, which avoid thermal cycling, are often confounded by primer dimers, off‐target priming, and other artifacts. Here, we show how a “self avoiding molecular recognition system” (SAMRS) eliminates these artifacts and gives clean amplicons in a helicase‐dependent isothermal amplification (SAMRS‐HDA). We also show that incorporating SAMRS into the 3′‐ends of primers facilitates the design and screening of primers for HDA assays. Finally, we show that SAMRS‐HDA can be twofold multiplexed, difficult to achieve with HDA using standard primers. Thus, SAMRS‐HDA is a more versatile approach than standard HDA, with a broader applicability for xNA‐targeted diagnostics and research.</div><div type="abstract" xml:lang="en">Primers containing self‐avoiding molecular recognition system (SAMRS) components can eliminate artifacts in helicase‐dependent isothermal amplification (HDA)—artifacts that are common in classical HDA that uses standard primers. SAMRS therefore greatly expands the versatility of HDA, and supports low levels of multiplexing.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Floride</li></region></list><tree><country name="États-Unis"><region name="Floride"><name sortKey="Yang, Zunyi" sort="Yang, Zunyi" uniqKey="Yang Z" first="Zunyi" last="Yang">Zunyi Yang</name></region><name sortKey="Benner, Steven A" sort="Benner, Steven A" uniqKey="Benner S" first="Steven A." last="Benner">Steven A. Benner</name><name sortKey="Benner, Steven A" sort="Benner, Steven A" uniqKey="Benner S" first="Steven A." last="Benner">Steven A. Benner</name><name sortKey="Benner, Steven A" sort="Benner, Steven A" uniqKey="Benner S" first="Steven A." last="Benner">Steven A. Benner</name><name sortKey="Bradley, Kevin M" sort="Bradley, Kevin M" uniqKey="Bradley K" first="Kevin M." last="Bradley">Kevin M. Bradley</name><name sortKey="Frye, Carole B" sort="Frye, Carole B" uniqKey="Frye C" first="Carole B." last="Frye">Carole B. Frye</name><name sortKey="Hoshika, Shuichi" sort="Hoshika, Shuichi" uniqKey="Hoshika S" first="Shuichi" last="Hoshika">Shuichi Hoshika</name><name sortKey="Hutter, Daniel" sort="Hutter, Daniel" uniqKey="Hutter D" first="Daniel" last="Hutter">Daniel Hutter</name><name sortKey="Hutter, Daniel" sort="Hutter, Daniel" uniqKey="Hutter D" first="Daniel" last="Hutter">Daniel Hutter</name><name sortKey="Mclendon, Chris" sort="Mclendon, Chris" uniqKey="Mclendon C" first="Chris" last="Mclendon">Chris Mclendon</name><name sortKey="Yang, Zunyi" sort="Yang, Zunyi" uniqKey="Yang Z" first="Zunyi" last="Yang">Zunyi Yang</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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