An approach to the use of stable isotopes for DNA sequencing

K. Bruce Jacobson; H. F. Arlinghaus; H. W. Schmitt; R. A. Sachleben; Gilbert M. Brown; N. Thonnard; F. V. Sloop; R. S. Foote; F. W. Larimer; R. P. Woychik; M. Wendy England; K. L. Burchett; Dan A. Jacobson

doi:10.1016/0888-7543(91)90220-9

An approach to the use of stable isotopes for DNA sequencing

K. Bruce Jacobson, H. F. Arlinghaus, H. W. Schmitt, R. A. Sachleben, Gilbert M. Brown, N. Thonnard, F. V. Sloop, R. S. Foote, F. W. Larimer, R. P. Woychik, M. Wendy England, K. L. Burchett, Dan A. Jacobson

Research output: Contribution to journal › Article › peer-review

27 Scopus citations

Abstract

The sequencing of DNA by current procedures involves the use of radioisotopic or fluorescent labels. We propose that stable isotopes can be used as such labels and that the large number of stable isotopes available would allow multiplexing so that many DNA segments could be sequenced simultaneously. We have developed methods to use ⁵⁷Fe₂O₃ to synthesize ferrocene and to attach the ferrocene to the 5′ end of oligonucleotides. The ⁵⁷Fe-labeled M13 universal primer functioned normally in a Sanger sequencing procedure. When a ⁵⁷Fe-labeled oligonucleotide had migrated on a polyacrylamide gel it was readily located on the dried gel by scanning with resonance ionization spectroscopy (RIS) coupled with mass spectrometry. Using a ⁵⁷Fe-labeled primer in a PCR reaction a 2000-bp DNA was produced that was detected by RIS on nylon membrane after agarose electrophoresis. The rapid analysis features of RIS coupled with the multispectral multiplexing possibilities of stable isotopes should significantly increase the rate of determination of DNA sequences.

Original language	English
Pages (from-to)	51-59
Number of pages	9
Journal	Genomics
Volume	9
Issue number	1
DOIs	https://doi.org/10.1016/0888-7543(91)90220-9
State	Published - Jan 1991
Externally published	Yes

Funding

This work was supported the Oak Ridge National by the Exploratory Studies Program of Laboratory and by the Office of Health ’ This research was sponsored by the Exploratory Studies Program of the Oak Ridge National Laboratory and the Office of Health and Environmental Research, U.S. Department of Energy, under Contract DE-AC05-84021400 with the Martin Marietta Energy Systems, Inc. The U.S. Government’s right to retain a nonexclusive royalty-free license in and to the copyright covering this paper, for governmental purposes, is acknowledged. ’ To whom correspondence and requests for reprints should be addressed. 3 Present address: Department of Pharmacology, University North Carolina School of Medicine, Chapel Hill, NC 27514.

Funders	Funder number
Environmental Research
Office of Health and Environmental Research
U.S. Department of Energy	DE-AC0584021400, DE-AC-05.89ER80735
Office of the National Coordinator for Health Information Technology
Oak Ridge National Laboratory

Access to Document

10.1016/0888-7543(91)90220-9

Cite this

@article{f5c6d7e2f0d54f2886dac06a25c535f8,

title = "An approach to the use of stable isotopes for DNA sequencing",

abstract = "The sequencing of DNA by current procedures involves the use of radioisotopic or fluorescent labels. We propose that stable isotopes can be used as such labels and that the large number of stable isotopes available would allow multiplexing so that many DNA segments could be sequenced simultaneously. We have developed methods to use 57Fe2O3 to synthesize ferrocene and to attach the ferrocene to the 5′ end of oligonucleotides. The 57Fe-labeled M13 universal primer functioned normally in a Sanger sequencing procedure. When a 57Fe-labeled oligonucleotide had migrated on a polyacrylamide gel it was readily located on the dried gel by scanning with resonance ionization spectroscopy (RIS) coupled with mass spectrometry. Using a 57Fe-labeled primer in a PCR reaction a 2000-bp DNA was produced that was detected by RIS on nylon membrane after agarose electrophoresis. The rapid analysis features of RIS coupled with the multispectral multiplexing possibilities of stable isotopes should significantly increase the rate of determination of DNA sequences.",

author = "Jacobson, {K. Bruce} and Arlinghaus, {H. F.} and Schmitt, {H. W.} and Sachleben, {R. A.} and Brown, {Gilbert M.} and N. Thonnard and Sloop, {F. V.} and Foote, {R. S.} and Larimer, {F. W.} and Woychik, {R. P.} and England, {M. Wendy} and Burchett, {K. L.} and Jacobson, {Dan A.}",

year = "1991",

month = jan,

doi = "10.1016/0888-7543(91)90220-9",

language = "English",

volume = "9",

pages = "51--59",

journal = "Genomics",

issn = "0888-7543",

publisher = "Elsevier",

number = "1",

}

TY - JOUR

T1 - An approach to the use of stable isotopes for DNA sequencing

AU - Jacobson, K. Bruce

AU - Arlinghaus, H. F.

AU - Schmitt, H. W.

AU - Sachleben, R. A.

AU - Brown, Gilbert M.

AU - Thonnard, N.

AU - Sloop, F. V.

AU - Foote, R. S.

AU - Larimer, F. W.

AU - Woychik, R. P.

AU - England, M. Wendy

AU - Burchett, K. L.

AU - Jacobson, Dan A.

PY - 1991/1

Y1 - 1991/1

N2 - The sequencing of DNA by current procedures involves the use of radioisotopic or fluorescent labels. We propose that stable isotopes can be used as such labels and that the large number of stable isotopes available would allow multiplexing so that many DNA segments could be sequenced simultaneously. We have developed methods to use 57Fe2O3 to synthesize ferrocene and to attach the ferrocene to the 5′ end of oligonucleotides. The 57Fe-labeled M13 universal primer functioned normally in a Sanger sequencing procedure. When a 57Fe-labeled oligonucleotide had migrated on a polyacrylamide gel it was readily located on the dried gel by scanning with resonance ionization spectroscopy (RIS) coupled with mass spectrometry. Using a 57Fe-labeled primer in a PCR reaction a 2000-bp DNA was produced that was detected by RIS on nylon membrane after agarose electrophoresis. The rapid analysis features of RIS coupled with the multispectral multiplexing possibilities of stable isotopes should significantly increase the rate of determination of DNA sequences.

AB - The sequencing of DNA by current procedures involves the use of radioisotopic or fluorescent labels. We propose that stable isotopes can be used as such labels and that the large number of stable isotopes available would allow multiplexing so that many DNA segments could be sequenced simultaneously. We have developed methods to use 57Fe2O3 to synthesize ferrocene and to attach the ferrocene to the 5′ end of oligonucleotides. The 57Fe-labeled M13 universal primer functioned normally in a Sanger sequencing procedure. When a 57Fe-labeled oligonucleotide had migrated on a polyacrylamide gel it was readily located on the dried gel by scanning with resonance ionization spectroscopy (RIS) coupled with mass spectrometry. Using a 57Fe-labeled primer in a PCR reaction a 2000-bp DNA was produced that was detected by RIS on nylon membrane after agarose electrophoresis. The rapid analysis features of RIS coupled with the multispectral multiplexing possibilities of stable isotopes should significantly increase the rate of determination of DNA sequences.

UR - http://www.scopus.com/inward/record.url?scp=0026063745&partnerID=8YFLogxK

U2 - 10.1016/0888-7543(91)90220-9

DO - 10.1016/0888-7543(91)90220-9

M3 - Article

C2 - 2004768

AN - SCOPUS:0026063745

SN - 0888-7543

VL - 9

SP - 51

EP - 59

JO - Genomics

JF - Genomics

IS - 1

ER -

An approach to the use of stable isotopes for DNA sequencing

Abstract

Funding

Access to Document

Other files and links

Fingerprint

Cite this