Isolation of YAC insert sequences by representational difference analysis

Mieke Schutte; Luis T. Da Costa; Christopher A. Moskaluk; Ester Rozenblum; Xiaoping Guan; Pieter J. De Jong; Michael Bittner; Paul S. Meltzer; Jeffrey M. Trent; Scott E. Kern

doi:10.1093/nar/23.20.4127

Isolation of YAC insert sequences by representational difference analysis

Mieke Schutte
, Luis T. Da Costa
, Christopher A. Moskaluk
, Ester Rozenblum
, Xiaoping Guan
, Pieter J. De Jong
, Michael Bittner
, Paul S. Meltzer
, Jeffrey M. Trent
, Scott E. Kern^*

^*Corresponding author for this work

Surgery

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

12 Scopus citations

Abstract

We present a method for the isolation of YAC insert sequences by representational difference analysis (RDA). To achieve maximal representation of the sequences, the amplicons were generated from a Mbol digestion product. RDA was performed using a 970 kb insert YAC clone. After two rounds of re-association and selective amplification 92% of the difference product represented sequences derived from the YAC insert. Twenty insert-specific sequence-tagged sites were readily defined. The difference product was also successfully used to isolate microsatellite markers, to identify clones from a human PAC library and as a chromosome painting probe in fluorescence in situ hybridization.

Original language	English
Pages (from-to)	4127-4133
Number of pages	7
Journal	Nucleic Acids Research
Volume	23
Issue number	20
DOIs	https://doi.org/10.1093/nar/23.20.4127
State	Published - 25 Oct 1995

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10.1093/nar/23.20.4127

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@article{28e41f9e11d5443c935ccdb5247bddb8,

title = "Isolation of YAC insert sequences by representational difference analysis",

abstract = "We present a method for the isolation of YAC insert sequences by representational difference analysis (RDA). To achieve maximal representation of the sequences, the amplicons were generated from a Mbol digestion product. RDA was performed using a 970 kb insert YAC clone. After two rounds of re-association and selective amplification 92\% of the difference product represented sequences derived from the YAC insert. Twenty insert-specific sequence-tagged sites were readily defined. The difference product was also successfully used to isolate microsatellite markers, to identify clones from a human PAC library and as a chromosome painting probe in fluorescence in situ hybridization.",

author = "Mieke Schutte and \{Da Costa\}, \{Luis T.\} and Moskaluk, \{Christopher A.\} and Ester Rozenblum and Xiaoping Guan and \{De Jong\}, \{Pieter J.\} and Michael Bittner and Meltzer, \{Paul S.\} and Trent, \{Jeffrey M.\} and Kern, \{Scott E.\}",

note = "Funding Information: This work was supported by NIH SPORE grant in Gastrointestinal Cancer CA62924, by the Dutch Cancer Society KWF (MS) and by JNICT Scholarship BD1508/91-ID (LT da C). XG and PJ de J are supported by the Cancer Center Support grant at Roswell Park Cancer Institute CA-16056, by Department of Energy grant DE-FG02-94ER61883 and by NIH grant 1R01RG01165-01. SEK is a McDonnell Foundation Scholar.",

year = "1995",

month = oct,

day = "25",

doi = "10.1093/nar/23.20.4127",

language = "English",

volume = "23",

pages = "4127--4133",

journal = "Nucleic Acids Research",

issn = "0305-1048",

publisher = "Oxford University Press",

number = "20",

}

TY - JOUR

T1 - Isolation of YAC insert sequences by representational difference analysis

AU - Schutte, Mieke

AU - Da Costa, Luis T.

AU - Moskaluk, Christopher A.

AU - Rozenblum, Ester

AU - Guan, Xiaoping

AU - De Jong, Pieter J.

AU - Bittner, Michael

AU - Meltzer, Paul S.

AU - Trent, Jeffrey M.

AU - Kern, Scott E.

N1 - Funding Information: This work was supported by NIH SPORE grant in Gastrointestinal Cancer CA62924, by the Dutch Cancer Society KWF (MS) and by JNICT Scholarship BD1508/91-ID (LT da C). XG and PJ de J are supported by the Cancer Center Support grant at Roswell Park Cancer Institute CA-16056, by Department of Energy grant DE-FG02-94ER61883 and by NIH grant 1R01RG01165-01. SEK is a McDonnell Foundation Scholar.

PY - 1995/10/25

Y1 - 1995/10/25

N2 - We present a method for the isolation of YAC insert sequences by representational difference analysis (RDA). To achieve maximal representation of the sequences, the amplicons were generated from a Mbol digestion product. RDA was performed using a 970 kb insert YAC clone. After two rounds of re-association and selective amplification 92% of the difference product represented sequences derived from the YAC insert. Twenty insert-specific sequence-tagged sites were readily defined. The difference product was also successfully used to isolate microsatellite markers, to identify clones from a human PAC library and as a chromosome painting probe in fluorescence in situ hybridization.

AB - We present a method for the isolation of YAC insert sequences by representational difference analysis (RDA). To achieve maximal representation of the sequences, the amplicons were generated from a Mbol digestion product. RDA was performed using a 970 kb insert YAC clone. After two rounds of re-association and selective amplification 92% of the difference product represented sequences derived from the YAC insert. Twenty insert-specific sequence-tagged sites were readily defined. The difference product was also successfully used to isolate microsatellite markers, to identify clones from a human PAC library and as a chromosome painting probe in fluorescence in situ hybridization.

UR - https://www.scopus.com/pages/publications/0028817940

U2 - 10.1093/nar/23.20.4127

DO - 10.1093/nar/23.20.4127

M3 - Article

C2 - 7479076

AN - SCOPUS:0028817940

SN - 0305-1048

VL - 23

SP - 4127

EP - 4133

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 20

ER -

Isolation of YAC insert sequences by representational difference analysis

Abstract

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