Molecular Biology

Genotyping

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View the protocol "Genotyping by Oligonucleotide Ligation Assay (OLA)" at BioSupplynet.com

Stuart J. Macdonald
Department of Ecology and Evolutionary Biology, University of Kansas, Lawrence, Kansas 67705, USA
Department of Molecular Biosciences, University of Kansas, Lawrence, Kansas 67705, USA

ABSTRACT
This protocol describes the oligonucleotide ligation assay (OLA), which uses a set of three oligonucleotides, in combination with a thermostable Taq DNA ligase enzyme, to discriminate single-nucleotide polymorphism (SNP) alleles. Sixteen-plex OLA genotyping reactions are carried out, and allele-specific OLA products are detected on membrane arrays using radiolabeled probes.

Products Available for this Protocol
Protocol Material Description Product # Product Name Add to Cart
Adenosine 5'-triphosphate (ATP) solution A6559 Adenosine 5'-triphosphate disodium salt solution
Triton X-100 T8787 Triton® X-100, for molecular biology
NaOH 655104 Sodium hydroxide, reagent grade, 97%, powder
dNTPs DNTP10 Deoxynucleotide Set, 10 mM
Herring sperm DNA D7290 Deoxyribonucleic acid from herring sperm, single-stranded for hybridization
β-NAD N8285 β-Nicotinamide adenine dinucleotide, ~98%, pkg of 10 mg (per vial)
Oligonucleotides Custom Oligos
SSC S8015 SSC Buffer, powder blend
SSPE S8140 SSPE Buffer, powder blend
SDS L4390 Sodium dodecyl sulfate, for molecular biology, ≥98.5% (GC)
T4 polynucleotide kinase P4390 Polynucleotide Kinase from T4-infected Escherichia coli, buffered aqueous glycerol solution
Tris-Cl T5941 Trizma® hydrochloride, for molecular biology, Biotechnology Performance Certified, cell culture tested, ≥99.0% (titration)
Taq DNA ligase D2886 DNA Ligase from T4-infected Escherichia coli, buffered aqueous glycerol solution
MgCl2 M4880 Magnesium chloride, insect cell culture tested
DTT D9779 DL-Dithiothreitol, for molecular biology, ≥98% (TLC), ≥99% (titration)
NaCl S6191 Sodium chloride, Biotechnology Performance Certified, ≥99.5% (titration), Cell Culture Tested
KCl P9541 Potassium chloride, for molecular biology, ≥99%
Taq DNA polymerase D1806 Taq DNA Polymerase from Thermus aquaticus, with 10× reaction buffer containing MgCl2, recombinant, expressed in Escherichia coli

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