Using synthetic oligonucleotides as probes.
The protocols in this unit describe procedures for using mixtures of 32P-labeled oligonucleotides to screen recombinant DNA clones bound to nitrocellulose filters. A partial amino acid sequence of a protein is used to predict the nucleotide sequence of the gene that would encode it. A mixture of oligonucleotides is chosen that includes all possible nucleotide sequences encoding that amino acid sequence. This mixture of oligonucleotides is then used to screen a recombinant DNA library for the corresponding clones. In some cases however, the exact nucleotide sequence of a desired clone is known and it is possible to use a unique oligonucleotide as a probe.
Duby
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A
The University of Texas Health Science Center at Dallas, Dallas, Texas, USA.
Jacobs
K A
KA
Celeste
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A
D001345:Autoradiography; D002951:Citrates; D007202:Indicators and Reagents; D007553:Isotope Labeling / Q000379:methods; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D010761:Phosphorus Radioisotopes; D000644:Quaternary Ammonium Compounds; D012965:Sodium Chloride; D000077559:Sodium Citrate; D013816:Thermodynamics
D002951:Citrates; D007202:Indicators and Reagents; D015345:Oligonucleotide Probes; D009841:Oligonucleotides; D010761:Phosphorus Radioisotopes; D000644:Quaternary Ammonium Compounds; D000077559:Sodium Citrate; D012965:Sodium Chloride; C027917:tetramethylammonium