Simple and Easy DNA Mapping Method Using Peptide Nucleic Acid (PNA) Tagging

被引：4

作者：

Kim, J. ^{[1
,2
]}

Park, J. H. ^{[1
]}

Lee, S. ^{[3
]}

Shin, H. J. ^{[2
]}

机构：

[1] Korea Univ, Sch Elect Engn, Seoul 136701, South Korea

[2] Korea Inst Sci & Technol, Ctr Bion, Seoul 136791, South Korea

[3] Korea Univ, Korea Inst Sci & Technol, Ctr Theragnosis, Seoul 136701, South Korea

来源：

JOURNAL OF NANOSCIENCE AND NANOTECHNOLOGY | 2014年 / 14卷 / 03期

关键词：

DNA Sequencing; DNA Mapping; DNA Stretching; PNA; Single Molecule Detection; SINGLE-MOLECULE DETECTION; GEL-ELECTROPHORESIS;

D O I：

10.1166/jnn.2014.8494

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

In this paper, we introduce a simple and easy DNA mapping method using peptide nucleic acid (PNA) on glass cover-slips based on optical detection. PNA and glass cover-slips were used as easy tagging and stretching method, separately. The PNA can be invaded lambda (lambda) DNA at the sequences we wished to tag without any additional materials. Alexa-488 fluorophore-conjugated PNA was designed to invade at four sites (sequence: AAGAAGAA) of lambda DNA with robust and exact binding. Fluorophore tagged lambda DNA was stretched in electrical layer coated glass cover-slips. The lambda DNA was stretched as 9.2 um in length. It is approximately 60% of theoretical length of lambda DNA and sufficient length to promise optical resolution. The sites of desired sequence tagged by PNA were well detected in stretched lambda DNA.

引用

页码：2477 / 2481

页数：5

共 11 条

[1] Rapid DNA sequencing of more than 1000 bases per run by capillary electrophoresis using replaceable linear polyacrylamide solutions [J].

Carrilho, E ;

RuizMartinez, MC ;

Berka, J ;

Smirnov, I ;

Goetzinger, W ;

Miller, AW ;

Brady, D ;

Karger, BL .

ANALYTICAL CHEMISTRY, 1996, 68 (19) :3305-3313

[2] DNA mapping using microfluidic stretching and single-molecule detection of fluorescent site-specific tags [J].

Chan, EY ;

Goncalves, NM ;

Haeusler, RA ;

Hatch, AJ ;

Larson, JW ;

Maletta, AM ;

Yantz, GR ;

Carstea, ED ;

Fuchs, M ;

Wong, GG ;

Gullans, SR ;

Gilmanshin, R .

GENOME RESEARCH, 2004, 14 (06) :1137-1146

[3] A simple DNA stretching method for fluorescence imaging of single DNA molecules [J].

Chan, Ting-Fung ;

Ha, Connie ;

Phong, Angie ;

Cai, Dongmei ;

Wan, Eunice ;

Leung, Lucinda ;

Kwok, Pui-Yan ;

Xiao, Ming .

NUCLEIC ACIDS RESEARCH, 2006, 34 (17)

[4] An experimental study of mechanism and specificity of peptide nucleic acid (PNA) binding to duplex DNA [J].

Kuhn, H ;

Demidov, VV ;

Nielsen, PE ;

Frank-Kamenetskii, MD .

JOURNAL OF MOLECULAR BIOLOGY, 1999, 286 (05) :1337-1345

[5] Double duplex invasion by peptide nucleic acid: A general principle for sequence-specific targeting of double-stranded DNA [J].

Lohse, J ;

Dahl, O ;

Nielsen, PE .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (21) :11804-11808

[6] OPTICAL MAPPING OF BACTERIOPHAGE-LAMBDA CLONES USING RESTRICTION ENDONUCLEASES [J].

MENG, X ;

BENSON, K ;

CHADA, K ;

HUFF, EJ ;

SCHWARTZ, DC .

NATURE GENETICS, 1995, 9 (04) :432-438

[7] RAPID RESTRICTION MAPPING OF DNA CLONED IN LAMBDA PHAGE VECTORS [J].

RACKWITZ, HR ;

ZEHETNER, G ;

FRISCHAUF, AM ;

LEHRACH, H .

GENE, 1984, 30 (1-3) :195-200

[8] DNA SEQUENCING WITH CHAIN-TERMINATING INHIBITORS [J].

SANGER, F ;

NICKLEN, S ;

COULSON, AR .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1977, 74 (12) :5463-5467

[9] DETECTION OF SPECIFIC SEQUENCES AMONG DNA FRAGMENTS SEPARATED BY GEL-ELECTROPHORESIS [J].

SOUTHERN, EM .

JOURNAL OF MOLECULAR BIOLOGY, 1975, 98 (03) :503-+

[10] CAPILLARY GEL-ELECTROPHORESIS FOR RAPID, HIGH-RESOLUTION DNA SEQUENCING [J].

SWERDLOW, H ;

GESTELAND, R .

NUCLEIC ACIDS RESEARCH, 1990, 18 (06) :1415-1419

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