|
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 66045, USA
Department of Molecular Biosciences, University of Kansas, Lawrence, Kansas 66045, 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 | |
back to top
View all available Cold Spring Harbor Laboratory Press Protocols
Product Association Disclaimer: The Sigma-Aldrich products listed for this specific protocol were selected either to match or to supplement the products listed within the actual protocol. The products/reagents from Sigma-Aldrich have been qualified for usage, but may not have been validated for this specific application. Please refer to the detailed product description on the usage of specific products of interest.
|
