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

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
Externally published	Yes

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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",

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 - http://www.scopus.com/inward/record.url?scp=0028817940&partnerID=8YFLogxK

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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