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The sample DNA is denatured, resulting in single-stranded sample DNA. Pairs of probes are hybridized to the sample DNA, with each probe pair designed to query for the presence of a particular DNA sequence. Ligase is applied to the hybridized DNA, combining probe pairs that are hybridized immediately next to each other into a single strand of ...
Multiple probe designs may be useful in identifying extraneous factors which may be influencing your results. Lastly, experimenters should avoid gathering data during sessions alone. If in-session data is gathered a note of the dates should be tagged to each measurement in order to provide an accurate time-line for potential reviewers.
In addition, with this design, bad probes affect all genotypes at a given locus equally. [3] For instance, since MIP probes can assay multiple genotypes at a particular genomic locus, if the probe for a given locus does not work (e.g. fails to properly hybridize to the genomic target), none of the genotypes at this locus will be detected.
In the oligonucleotide ligase assay, two probes are designed; an allele-specific probe which hybridizes to the target DNA so that its 3' base is situated directly over the SNP nucleotide and a second probe that hybridizes the template upstream (downstream in the complementary strand) of the SNP polymorphic site providing a 5' end for the ...
Conventional SNP typing methods are typically time-consuming and expensive, requiring several probe based assays to be multiplexed together or the use of DNA microarrays. HRM is more cost-effective and reduces the need to design multiple pairs of primers and the need to purchase expensive probes.
Multiplex-PCR consists of multiple primer sets within a single PCR mixture to produce amplicons of varying sizes that are specific to different DNA sequences. By targeting multiple sequences at once, additional information may be gained from a single test run that otherwise would require several times the reagents and more time to perform.