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Title: Preparation of 13C/15N-labeled oligomers using the polymerase chain reaction

Abstract

Preparation of .sup.13 C/.sup.15 N-labeled DNA oligomers using the polymerase chain reaction (PCR). A PCR based method for uniform (.sup.13 C/.sup.15 N)-labeling of DNA duplexes is described. Multiple copies of a blunt-ended duplex are cloned into a plasmid, each copy containing the sequence of interest and restriction Hinc II sequences at both the 5' and 3' ends. PCR using bi-directional primers and uniformly .sup.13 C/.sup.15 N-labeled dNTP precursors generates labeled DNA duplexes containing multiple copies of the sequence of interest. Twenty-four cycles of PCR, followed by restriction and purification, gave the uniformly .sup.13 C/.sup.15 N-labeled duplex sequence with a 30% yield. Such labeled duplexes find significant applications in multinuclear magnetic resonance spectroscopy.

Inventors:
 [1];  [2];  [2]
  1. (Los Alamos, NM)
  2. (Santa Fe, NM)
Issue Date:
Research Org.:
Los Alamos National Laboratory (LANL), Los Alamos, NM
OSTI Identifier:
873843
Patent Number(s):
6258567
Assignee:
Regents of University of California (Los Alamos, NM) LANL
DOE Contract Number:  
W-7405-ENG-36
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
preparation; 13c; 15n-labeled; oligomers; polymerase; chain; reaction; 13; 15; n-labeled; dna; pcr; based; method; uniform; -labeling; duplexes; described; multiple; copies; blunt-ended; duplex; cloned; plasmid; copy; containing; sequence; restriction; hinc; ii; sequences; bi-directional; primers; uniformly; dntp; precursors; generates; labeled; twenty-four; cycles; followed; purification; 30; yield; significant; applications; multinuclear; magnetic; resonance; spectroscopy; resonance spectroscopy; labeled dna; chain reaction; magnetic resonance; polymerase chain; nuclear magnetic; based method; containing multiple; /435/999/

Citation Formats

Chen, Xian, Gupta, Goutam, and Bradbury, E. Morton. Preparation of 13C/15N-labeled oligomers using the polymerase chain reaction. United States: N. p., 2001. Web.
Chen, Xian, Gupta, Goutam, & Bradbury, E. Morton. Preparation of 13C/15N-labeled oligomers using the polymerase chain reaction. United States.
Chen, Xian, Gupta, Goutam, and Bradbury, E. Morton. Mon . "Preparation of 13C/15N-labeled oligomers using the polymerase chain reaction". United States. https://www.osti.gov/servlets/purl/873843.
@article{osti_873843,
title = {Preparation of 13C/15N-labeled oligomers using the polymerase chain reaction},
author = {Chen, Xian and Gupta, Goutam and Bradbury, E. Morton},
abstractNote = {Preparation of .sup.13 C/.sup.15 N-labeled DNA oligomers using the polymerase chain reaction (PCR). A PCR based method for uniform (.sup.13 C/.sup.15 N)-labeling of DNA duplexes is described. Multiple copies of a blunt-ended duplex are cloned into a plasmid, each copy containing the sequence of interest and restriction Hinc II sequences at both the 5' and 3' ends. PCR using bi-directional primers and uniformly .sup.13 C/.sup.15 N-labeled dNTP precursors generates labeled DNA duplexes containing multiple copies of the sequence of interest. Twenty-four cycles of PCR, followed by restriction and purification, gave the uniformly .sup.13 C/.sup.15 N-labeled duplex sequence with a 30% yield. Such labeled duplexes find significant applications in multinuclear magnetic resonance spectroscopy.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2001},
month = {1}
}

Patent:

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