Document Detail


Variability of the random amplified polymorphic DNA assay among thermal cyclers, and effects of primer and DNA concentration.
MedLine Citation:
PMID:  8232346     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The reproducibility of the generation of random amplified polymorphic DNA fragments from three commonly used thermal cyclers was determined using identical assay conditions. In all cases, different results were obtained from the three instruments. Variation in the length of the primer (20 nt or 10 nt) did not have any effect on the reproducibility of the assays from the three machines tested. A DNA concentration of 1 ng generated poorly staining DNA fragments whereas concentrations between 10 ng and 100 ng gave similar banding patterns when using the same thermal cycler. Low concentrations of primer (0.05 microM) did not produce any detectable DNA fragments. Increased primer concentrations of 0.25 microM or higher generated intensely staining DNA fragments, and concentrations above 0.5 microM did not improve the clarity of the banding patterns but did direct the synthesis of increasing amounts of very short DNA fragments. Surprisingly, the 20 nt-long primer was able to direct the synthesis of more DNA fragments than a primer of only 10 nt long.
Authors:
J M MacPherson; P E Eckstein; G J Scoles; A A Gajadhar
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Publication Detail:
Type:  Comparative Study; Journal Article    
Journal Detail:
Title:  Molecular and cellular probes     Volume:  7     ISSN:  0890-8508     ISO Abbreviation:  Mol. Cell. Probes     Publication Date:  1993 Aug 
Date Detail:
Created Date:  1993-12-07     Completed Date:  1993-12-07     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8709751     Medline TA:  Mol Cell Probes     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  293-9     Citation Subset:  IM    
Affiliation:
Health of Animals Laboratory, Agriculture Canada, Saskatoon, Saskatchewan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
DNA, Protozoan / genetics
Molecular Sequence Data
Polymerase Chain Reaction / instrumentation*,  methods*
RNA, Ribosomal, 18S / genetics
Toxoplasma / genetics
Chemical
Reg. No./Substance:
0/DNA, Protozoan; 0/RNA, Ribosomal, 18S

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


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