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28 Cards in this Set
- Front
- Back
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What does the southern blot test?
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It is the classic method for DNA analysis. DNA is broken down by enzymes to produce fragments, the fragments are then separated by electrophoresis and transferred to a solid support medium, then a labeled probe is added and binds to the complementary base sequence forming a band.
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What are three solid phase assays?
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capture hybridization, southern blot, northern blot.
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How is RNA different from DNA?
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It is single stranded, contains a uracil base instead of thymine, the sugar is ribose instead of deoxyribose.
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What can nucleic acid techniques detect?
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Infectious organisms, gene rearrangements, changes in tumor suppressor factors, chromosomal breaks or changes, prenatal diagnosis of inherited disease or carrier status, identity or nonidentity, available tissue donors.
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What does the Western blot test for?
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Detects specific antibodies, confirmation test for positive serologic testing of HIV.
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State the steps of DNA replication.
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DNA is denatured, the single-stranded DNA serves as a template for the mRNA, mRNA is released from the template DNA, DNA reanneals.
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Name some nucleic acid probe techniques.
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Solid Phase Assays( capture hybridization, southern blot, northern blot); Solution Based Assays( protection assays, hybrid capture assays); In situ hybridiztion assays (in tissue)
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What do nucleic acid techniques target?
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DNA and RNA
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What are three uses for hybridization?
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Identifying DNA in a crime scene, to determine who is related, and to see what percentage of similarity to species have
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What does the northern blot test?
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It tests for RNA, same procedure and southern but with RNA.
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What are two solution based assays?
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protection assays, hybrid capture assays.
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What is the purpose of the modified dot-blot?
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Designed to overcome background problems with use of unpure sample
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What does the southern blot extract?
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DNA
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What are three probe types used in amplification?
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Nucleic acids, Cloned DNA or RNA, or synthetic DNA
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What does the Northern blot extract?
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RNA
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Explain Dot-Blot hybridization.
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clinical samples are directly applied to membrane surface, membrane is heated to denature DNA, labeled probe is added(qualitatively detects target sequence)
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What is the sandwich assay also known as?
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Modified Dot-blot
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What are some amplification techniques and examples?
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Nucleic Acid Amplification( PCR, nucleic acid-based sequence amplification, transcription-mediated amplification.) :Probe Amplification ( ligase chain reaction): Signal Amplification ( branched-chain DNA Assay)
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What are the functions of the two probes used in modified dot blot?
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1st is bound to membrane and captures target sample DNA, 2nd binds to different site on target DNA and has label for detection.
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What is the gold standard for molecular application?
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DNA sequencing
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What is the principle and purpose of RFLP?
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mutated DNA can alter the length of the DNA fragment and the purpose is it evaluates differences in genomic DNA. It can establish identity or nonidentity, and identify genes associated with disease
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What are four uses for detecting point mutation?
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leukemia classification, molecular staging of tumors, characterization of microbial agents, determining genes associated with such things as drug resistance in HIV.
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What is an example of signal amplification?
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Branched chain signal amplification (bDNA)
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What media is used in chip technology?
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microarrays, biochips
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What is the benefit of using chip technology?
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allows thousands of reactions to be performed at once
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Discuss PCR
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amplifies target DNA through a series of amplification cycles, can amplify target DNA sequence prior to RFLP, sequences are know as amplicons
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What does rt-PCR use as a target sequence?
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mRNA
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What is an example of probe amplification?
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Ligase Chain Reaction (LCR)
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