Construction of a DNA library from chromosome 4 of rice (Oryza sativa) by microdissection

Ying Wei Mao, Si Yuan Liang, Wen Qin Song, Xiu Lan Li, Rui Yang Chen

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

A simple method to create a chromosome-specific DNA library of rice, including microdissection, amplification, characterization and cloning, is described. Rice chromosome 4 from a metaphase cell has been isolated and amplified by the Linker Adapter PCR (LA-PCR). The PCR products were labeled as probes with DIG-11-dUTP using the random priming method. Southern blot analysis with rice genomic DNA and specific RFLP markers demonstrated that the PCR products were derived from rice chromosome 4. A large library comprising over 100,000 recombinant plasmid microclones from rice chromosome 4 was constructed. Colony hybridization showed that 58% of the clones contained single or low-copy sequences and 42% contained repetitive sequences. The size of inserts generated by PCR ranged from 140bp to 500bp.This method will facilitate cloning of the specific chromosome DNA markers and important genes of rice.

Original languageEnglish (US)
Pages (from-to)285-293
Number of pages9
JournalCell Research
Volume8
Issue number4
DOIs
StatePublished - Jan 1 1998

Fingerprint

Microdissection
Chromosomes, Human, Pair 4
Gene Library
Polymerase Chain Reaction
Genetic Markers
Organism Cloning
Nucleic Acid Repetitive Sequences
Metaphase
Southern Blotting
Restriction Fragment Length Polymorphisms
Libraries
Oryza
Plasmids
Clone Cells
Chromosomes
DNA
Genes

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

Mao, Ying Wei ; Liang, Si Yuan ; Song, Wen Qin ; Li, Xiu Lan ; Chen, Rui Yang. / Construction of a DNA library from chromosome 4 of rice (Oryza sativa) by microdissection. In: Cell Research. 1998 ; Vol. 8, No. 4. pp. 285-293.
@article{19cbcac95d424ec1943151b122591b19,
title = "Construction of a DNA library from chromosome 4 of rice (Oryza sativa) by microdissection",
abstract = "A simple method to create a chromosome-specific DNA library of rice, including microdissection, amplification, characterization and cloning, is described. Rice chromosome 4 from a metaphase cell has been isolated and amplified by the Linker Adapter PCR (LA-PCR). The PCR products were labeled as probes with DIG-11-dUTP using the random priming method. Southern blot analysis with rice genomic DNA and specific RFLP markers demonstrated that the PCR products were derived from rice chromosome 4. A large library comprising over 100,000 recombinant plasmid microclones from rice chromosome 4 was constructed. Colony hybridization showed that 58{\%} of the clones contained single or low-copy sequences and 42{\%} contained repetitive sequences. The size of inserts generated by PCR ranged from 140bp to 500bp.This method will facilitate cloning of the specific chromosome DNA markers and important genes of rice.",
author = "Mao, {Ying Wei} and Liang, {Si Yuan} and Song, {Wen Qin} and Li, {Xiu Lan} and Chen, {Rui Yang}",
year = "1998",
month = "1",
day = "1",
doi = "10.1038/cr.1998.28",
language = "English (US)",
volume = "8",
pages = "285--293",
journal = "Cell Research",
issn = "1001-0602",
publisher = "Nature Publishing Group",
number = "4",

}

Construction of a DNA library from chromosome 4 of rice (Oryza sativa) by microdissection. / Mao, Ying Wei; Liang, Si Yuan; Song, Wen Qin; Li, Xiu Lan; Chen, Rui Yang.

In: Cell Research, Vol. 8, No. 4, 01.01.1998, p. 285-293.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Construction of a DNA library from chromosome 4 of rice (Oryza sativa) by microdissection

AU - Mao, Ying Wei

AU - Liang, Si Yuan

AU - Song, Wen Qin

AU - Li, Xiu Lan

AU - Chen, Rui Yang

PY - 1998/1/1

Y1 - 1998/1/1

N2 - A simple method to create a chromosome-specific DNA library of rice, including microdissection, amplification, characterization and cloning, is described. Rice chromosome 4 from a metaphase cell has been isolated and amplified by the Linker Adapter PCR (LA-PCR). The PCR products were labeled as probes with DIG-11-dUTP using the random priming method. Southern blot analysis with rice genomic DNA and specific RFLP markers demonstrated that the PCR products were derived from rice chromosome 4. A large library comprising over 100,000 recombinant plasmid microclones from rice chromosome 4 was constructed. Colony hybridization showed that 58% of the clones contained single or low-copy sequences and 42% contained repetitive sequences. The size of inserts generated by PCR ranged from 140bp to 500bp.This method will facilitate cloning of the specific chromosome DNA markers and important genes of rice.

AB - A simple method to create a chromosome-specific DNA library of rice, including microdissection, amplification, characterization and cloning, is described. Rice chromosome 4 from a metaphase cell has been isolated and amplified by the Linker Adapter PCR (LA-PCR). The PCR products were labeled as probes with DIG-11-dUTP using the random priming method. Southern blot analysis with rice genomic DNA and specific RFLP markers demonstrated that the PCR products were derived from rice chromosome 4. A large library comprising over 100,000 recombinant plasmid microclones from rice chromosome 4 was constructed. Colony hybridization showed that 58% of the clones contained single or low-copy sequences and 42% contained repetitive sequences. The size of inserts generated by PCR ranged from 140bp to 500bp.This method will facilitate cloning of the specific chromosome DNA markers and important genes of rice.

UR - http://www.scopus.com/inward/record.url?scp=0032241844&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032241844&partnerID=8YFLogxK

U2 - 10.1038/cr.1998.28

DO - 10.1038/cr.1998.28

M3 - Article

C2 - 9934536

AN - SCOPUS:0032241844

VL - 8

SP - 285

EP - 293

JO - Cell Research

JF - Cell Research

SN - 1001-0602

IS - 4

ER -