Document Detail


Immiscible phase nucleic acid purification eliminates PCR inhibitors with a single pass of paramagnetic particles through a hydrophobic liquid.
MedLine Citation:
PMID:  20581047     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Extraction and purification of nucleic acids from complex biological samples for PCR are critical steps because inhibitors must be removed that can affect reaction efficiency and the accuracy of results. This preanalytical processing generally involves capturing nucleic acids on microparticles that are then washed with a series of buffers to desorb and dilute out interfering substances. We have developed a novel purification method that replaces multiple wash steps with a single pass of paramagnetic particles (PMPs) though an immiscible hydrophobic liquid. Only two aqueous solutions are required: a lysis buffer, in which nucleic acids are captured on PMPs, and an elution buffer, in which they are released for amplification. The PMPs containing the nucleic acids are magnetically transported through a channel containing liquid wax that connects the lysis chamber to the elution chamber in a specially designed cartridge. Transporting PMPs through the immiscible phase yielded DNA and RNA as pure as that obtained after extensive wash steps required by comparable purification methods. Our immiscible-phase process has been applied to targets in whole blood, plasma, and urine and will enable the development of faster and simpler purification systems.
Authors:
Kunal Sur; Sally M McFall; Emilie T Yeh; Sujit R Jangam; Mark A Hayden; Stephen D Stroupe; David M Kelso
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-06-25
Journal Detail:
Title:  The Journal of molecular diagnostics : JMD     Volume:  12     ISSN:  1943-7811     ISO Abbreviation:  J Mol Diagn     Publication Date:  2010 Sep 
Date Detail:
Created Date:  2010-08-31     Completed Date:  2011-01-13     Revised Date:  2011-09-13    
Medline Journal Info:
Nlm Unique ID:  100893612     Medline TA:  J Mol Diagn     Country:  United States    
Other Details:
Languages:  eng     Pagination:  620-8     Citation Subset:  IM    
Affiliation:
Center for Innovation in Global Health Technologies, Northwestern University, Evanston, Illinois 60208, USA.
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MeSH Terms
Descriptor/Qualifier:
Chlamydia Infections / diagnosis,  microbiology
Chlamydia trachomatis / genetics,  isolation & purification
Gonorrhea / diagnosis
HIV Infections / diagnosis
HIV-1 / genetics,  isolation & purification
Humans
Neisseria gonorrhoeae / genetics,  isolation & purification
Nucleic Acids / isolation & purification*
Reverse Transcriptase Polymerase Chain Reaction*
Chemical
Reg. No./Substance:
0/Nucleic Acids
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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