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Title: Methods for producing partially digested restriction DNA fragments and for producing a partially modified PCR product

Abstract

The present invention is an improved method of making a partially modified PCR product from a DNA fragment with a polymerase chain reaction (PCR). In a standard PCR process, the DNA fragment is combined with starting deoxynucleoside triphosphates, a primer, a buffer and a DNA polymerase in a PCR mixture. The PCR mixture is then reacted in the PCR producing copies of the DNA fragment. The improvement of the present invention is adding an amount of a modifier at any step prior to completion of the PCR process thereby randomly and partially modifying the copies of the DNA fragment as a partially modified PCR product. The partially modified PCR product may then be digested with an enzyme that cuts the partially modified PCR product at unmodified sites thereby producing an array of DNA restriction fragments.

Inventors:
 [1]
  1. Richland, WA
Issue Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
OSTI Identifier:
872938
Patent Number(s):
6046039
Assignee:
Battelle Memorial Institute (Richland, WA)
Patent Classifications (CPCs):
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
C - CHEMISTRY C12 - BIOCHEMISTRY C12P - FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {
DOE Contract Number:  
AC06-76RL01830
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
methods; producing; partially; digested; restriction; dna; fragments; modified; pcr; product; improved; method; fragment; polymerase; chain; reaction; standard; process; combined; starting; deoxynucleoside; triphosphates; primer; buffer; mixture; reacted; copies; improvement; adding; amount; modifier; step; prior; completion; randomly; modifying; enzyme; cuts; unmodified; sites; array; pcr product; partially modified; modified pcr; chain reaction; dna polymerase; improved method; dna fragments; polymerase chain; dna fragment; restriction fragments; restriction fragment; nucleoside triphosphate; /435/436/999/

Citation Formats

Wong, Kwong-Kwok. Methods for producing partially digested restriction DNA fragments and for producing a partially modified PCR product. United States: N. p., 2000. Web.
Wong, Kwong-Kwok. Methods for producing partially digested restriction DNA fragments and for producing a partially modified PCR product. United States.
Wong, Kwong-Kwok. Sat . "Methods for producing partially digested restriction DNA fragments and for producing a partially modified PCR product". United States. https://www.osti.gov/servlets/purl/872938.
@article{osti_872938,
title = {Methods for producing partially digested restriction DNA fragments and for producing a partially modified PCR product},
author = {Wong, Kwong-Kwok},
abstractNote = {The present invention is an improved method of making a partially modified PCR product from a DNA fragment with a polymerase chain reaction (PCR). In a standard PCR process, the DNA fragment is combined with starting deoxynucleoside triphosphates, a primer, a buffer and a DNA polymerase in a PCR mixture. The PCR mixture is then reacted in the PCR producing copies of the DNA fragment. The improvement of the present invention is adding an amount of a modifier at any step prior to completion of the PCR process thereby randomly and partially modifying the copies of the DNA fragment as a partially modified PCR product. The partially modified PCR product may then be digested with an enzyme that cuts the partially modified PCR product at unmodified sites thereby producing an array of DNA restriction fragments.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2000},
month = {1}
}

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Works referenced in this record:

Control of partial digestion combining the enzymes dam methylase and MboI
journal, January 1989


PCR with 5-methyl-dCTP replacing dCTP
journal, January 1991


New cloning vectors and techniques for easy and rapid restriction mapping
journal, July 1988