Document Detail


Evaluation of DNA probe removal from nylon membrane.
MedLine Citation:
PMID:  1362069     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Genetic fingerprinting is one of the most challenging applications of any hybridization membranes. Forensic DNA fingerprinting typically uses samples in the range of 100-400 ng of genomic DNA. To ensure the ability to successfully reprobe the samples, it is imperative that repeated stripping of sample DNA be minimized while stripping of the probe DNA be maximized. By using standard dilutions of K562 cell line, we compared the following three stripping techniques: NaOH at 25 degrees C, formamide (HCONH2) at 65 degrees C and 0.1 x standard saline citrate and 0.1% sodium dodecyl sulfate at 95 degrees C (high-temperature stripping). The largest amount of genomic DNA was stripped from the membrane with NaOH, with the other two techniques removing less. Formamide and high-temperature procedures resulted in a loss of approximately 5-10 ng of DNA per strip. In contrast, the NaOH resulted in a loss of approximately 10-20 ng per strip.
Authors:
K Noppinger; G Duncan; D Ferraro; S Watson; J Ban
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  BioTechniques     Volume:  13     ISSN:  0736-6205     ISO Abbreviation:  BioTechniques     Publication Date:  1992 Oct 
Date Detail:
Created Date:  1993-02-08     Completed Date:  1993-02-08     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  8306785     Medline TA:  Biotechniques     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  572-5     Citation Subset:  IM    
Affiliation:
Broward County Sheriff's Crime Laboratory, DNA Unit, Ft. Lauderdale, FL 33301.
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MeSH Terms
Descriptor/Qualifier:
Autoradiography
DNA Fingerprinting / methods*
DNA Probes / isolation & purification*
Evaluation Studies as Topic
Formamides
Hot Temperature
Humans
Membranes, Artificial
Nucleic Acid Hybridization
Nylons*
Polymorphism, Restriction Fragment Length
Sodium Hydroxide
Chemical
Reg. No./Substance:
0/DNA Probes; 0/Formamides; 0/Membranes, Artificial; 0/Nylons; 1310-73-2/Sodium Hydroxide; 75-12-7/formamide

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


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