Preparation of 13C/15N-labeled oligomers using the polymerase chain reaction
- Los Alamos, NM
- Santa Fe, NM
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.
- Research Organization:
- Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)
- DOE Contract Number:
- W-7405-ENG-36
- Assignee:
- Regents of University of California (Los Alamos, NM)
- Patent Number(s):
- US 6258567
- OSTI ID:
- 873843
- Country of Publication:
- United States
- Language:
- English
In vivo localization of DNA sequences and visualization of large-scale chromatin organization using lac operator/repressor recognition
|
journal | December 1996 |
NMR of enzymatically synthesized uniformly 13C15N-labeled DNA oligonucleotides.
|
journal | April 1995 |
Preparation of Uniformly Isotope-labeled DNA Oligonucleotides for NMR Spectroscopy
|
journal | January 1998 |
Similar Records
Kinetic mechanism of DNA polymerase I (Klenow)
Two-dimensional NMR studies of staphylococcal nuclease. 2. Sequence-specific assignments of carbon-13 and nitrogen-15 signals from the nuclease H124L-thymidine 3 prime ,5 prime -bisphosphate-Ca sup 2+ ternary complex
Related Subjects
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/