Genetic Mapping of Soybean [Glycine max (L.) Merr.] Using Random Amplified Polymorphic DNA (RAPD)

Arnaldo Ribeiro Ferreira, Paul S Keim

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Random amplified polymorphic DNA (RAPD) is based on DNA amplification by polymerase chain reaction (PCR) of random DNA segments using single arbitrary nucleotide sequences. We have adapted the assay to soybeans by using Stoffel Fragment DNA polymerase and by optimizing the reaction conditions. To increase the percentage of RAPD polymorphisms, the DNA template was digested with restriction enzymes before amplification. The combination of twenty-four primers and five DNA template treatments (Undigested, DraI, EcoRI, HindIII, and TaqI digested) revealed 94 polymorphic DNA fragments differing between soybean lines PI437654 and BSR101. Many polymorphic DNA bands were found unreliable or non-scoreable after re-screening of primers and verification of marker-allele segregation with 20 recombinant inbred lines (RILs). However, 28 RAPD markers were consistently polymorphic between the parental lines and followed Mendelian expectations. The use of DNA templates digested with DraI, EcoRI, HindIII or TaqI increased three times the number of RAPD markers compared to undigested DNA template alone. The 28 RAPD markers obtained were further screened with 72 RILs and placed on an existing RFLP map.

Original languageEnglish (US)
Pages (from-to)335-354
Number of pages20
JournalPlant Molecular Biology Reporter
Volume15
Issue number4
StatePublished - 1997

Fingerprint

random amplified polymorphic DNA technique
Soybeans
Glycine
chromosome mapping
Glycine max
soybeans
DNA
Genetic Markers
DNA-Directed DNA Polymerase
inbred lines
genetic markers
gene segregation
DNA Primers
Amplification
DNA-directed DNA polymerase
Restriction Fragment Length Polymorphisms
restriction fragment length polymorphism
polymerase chain reaction
Polymerase chain reaction
Alleles

Keywords

  • Linkage mapping
  • PCR
  • RAPD
  • Restriction enzymes
  • Soybean

ASJC Scopus subject areas

  • Plant Science
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Genetic Mapping of Soybean [Glycine max (L.) Merr.] Using Random Amplified Polymorphic DNA (RAPD). / Ferreira, Arnaldo Ribeiro; Keim, Paul S.

In: Plant Molecular Biology Reporter, Vol. 15, No. 4, 1997, p. 335-354.

Research output: Contribution to journalArticle

@article{ecb3ff10a1704c76bedc1ece031c75c1,
title = "Genetic Mapping of Soybean [Glycine max (L.) Merr.] Using Random Amplified Polymorphic DNA (RAPD)",
abstract = "Random amplified polymorphic DNA (RAPD) is based on DNA amplification by polymerase chain reaction (PCR) of random DNA segments using single arbitrary nucleotide sequences. We have adapted the assay to soybeans by using Stoffel Fragment DNA polymerase and by optimizing the reaction conditions. To increase the percentage of RAPD polymorphisms, the DNA template was digested with restriction enzymes before amplification. The combination of twenty-four primers and five DNA template treatments (Undigested, DraI, EcoRI, HindIII, and TaqI digested) revealed 94 polymorphic DNA fragments differing between soybean lines PI437654 and BSR101. Many polymorphic DNA bands were found unreliable or non-scoreable after re-screening of primers and verification of marker-allele segregation with 20 recombinant inbred lines (RILs). However, 28 RAPD markers were consistently polymorphic between the parental lines and followed Mendelian expectations. The use of DNA templates digested with DraI, EcoRI, HindIII or TaqI increased three times the number of RAPD markers compared to undigested DNA template alone. The 28 RAPD markers obtained were further screened with 72 RILs and placed on an existing RFLP map.",
keywords = "Linkage mapping, PCR, RAPD, Restriction enzymes, Soybean",
author = "Ferreira, {Arnaldo Ribeiro} and Keim, {Paul S}",
year = "1997",
language = "English (US)",
volume = "15",
pages = "335--354",
journal = "Plant Molecular Biology Reporter",
issn = "0735-9640",
publisher = "Springer New York",
number = "4",

}

TY - JOUR

T1 - Genetic Mapping of Soybean [Glycine max (L.) Merr.] Using Random Amplified Polymorphic DNA (RAPD)

AU - Ferreira, Arnaldo Ribeiro

AU - Keim, Paul S

PY - 1997

Y1 - 1997

N2 - Random amplified polymorphic DNA (RAPD) is based on DNA amplification by polymerase chain reaction (PCR) of random DNA segments using single arbitrary nucleotide sequences. We have adapted the assay to soybeans by using Stoffel Fragment DNA polymerase and by optimizing the reaction conditions. To increase the percentage of RAPD polymorphisms, the DNA template was digested with restriction enzymes before amplification. The combination of twenty-four primers and five DNA template treatments (Undigested, DraI, EcoRI, HindIII, and TaqI digested) revealed 94 polymorphic DNA fragments differing between soybean lines PI437654 and BSR101. Many polymorphic DNA bands were found unreliable or non-scoreable after re-screening of primers and verification of marker-allele segregation with 20 recombinant inbred lines (RILs). However, 28 RAPD markers were consistently polymorphic between the parental lines and followed Mendelian expectations. The use of DNA templates digested with DraI, EcoRI, HindIII or TaqI increased three times the number of RAPD markers compared to undigested DNA template alone. The 28 RAPD markers obtained were further screened with 72 RILs and placed on an existing RFLP map.

AB - Random amplified polymorphic DNA (RAPD) is based on DNA amplification by polymerase chain reaction (PCR) of random DNA segments using single arbitrary nucleotide sequences. We have adapted the assay to soybeans by using Stoffel Fragment DNA polymerase and by optimizing the reaction conditions. To increase the percentage of RAPD polymorphisms, the DNA template was digested with restriction enzymes before amplification. The combination of twenty-four primers and five DNA template treatments (Undigested, DraI, EcoRI, HindIII, and TaqI digested) revealed 94 polymorphic DNA fragments differing between soybean lines PI437654 and BSR101. Many polymorphic DNA bands were found unreliable or non-scoreable after re-screening of primers and verification of marker-allele segregation with 20 recombinant inbred lines (RILs). However, 28 RAPD markers were consistently polymorphic between the parental lines and followed Mendelian expectations. The use of DNA templates digested with DraI, EcoRI, HindIII or TaqI increased three times the number of RAPD markers compared to undigested DNA template alone. The 28 RAPD markers obtained were further screened with 72 RILs and placed on an existing RFLP map.

KW - Linkage mapping

KW - PCR

KW - RAPD

KW - Restriction enzymes

KW - Soybean

UR - http://www.scopus.com/inward/record.url?scp=0031293031&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031293031&partnerID=8YFLogxK

M3 - Article

VL - 15

SP - 335

EP - 354

JO - Plant Molecular Biology Reporter

JF - Plant Molecular Biology Reporter

SN - 0735-9640

IS - 4

ER -