Polymerase Chain Reaction continued What are three important application of PCR? Amplification of DNA Identification of a target sequence Synthesis of a labeled antisense probe What is considered to be the method DNA Sequencing “Gold Standard” to which other molecular methods are compared? What hybridization methods are used for the Dot blot and reverse dot blot detection of disorder in which the DNA sequence of the mutated region has been identified (sickle cell) This hybridization method is used to detect Dot Blot base mutations using allele-specific oligonucleotides (ASOs) Unlike other assays, the dot blot does not require Enzyme digestion which two parts of the procedure? Electrophoretic separation of DNA fragments What are the basic steps of the procedure? Labeled oligonucleotide probes of 15-19 pairs are added. DNA is amplified in the region of the known mutations, denatured and applied to areas of membrane or filter. What is a Reverse Dot Blot? Variation of Dot Blot procedure How is the Reverse Dot Blot different? The ASO probes are bound to a filter and denatured DNA from the patient is added to the immobilized ASO. Hybridization occurs when base sequences are 100% complementary to the probe What is an example of a southern blot? This procedure is used for analysis of the proximal Northern Blot Blotting Protocol product of gene expression, mRNA. Cloned DNA probes can determine whether a given gene is expressed and, of so, how vigorously. This is a tissue-based molecular diagnostic assay. In Situ Hybridization Common in situ hybridization (ISH), including fluorescent ISH (FISH), probes are used in the diagnosis of hematological malignancies in clouding CMLA, AML, Burkitt lymphoma and other lymphomas. This gene expression profiling uses Microarrays _____ analysis. A surface (in this example, a glass microscope slide) contains thousands of spots. Each spot contains a large number of DNA fragments. For each spot, the DNA fragments are derived from one
Polymerase Chain Reaction continued What are three important application of PCR? Amplification of DNA Identification of a target sequence Synthesis of a labeled antisense probe What is considered to be the “Gold Standard” method to which other molecular methods are compared? DNA Sequencing What hybridization methods are used for the detection of disorder in which the DNA sequence of the mutated region has been identified (sickle cell) Dot blot and reverse dot blot This hybridization method is used to detect mutations using allele-specific oligonucleotides (ASOs) Dot Blot Unlike other assays, the dot blot does not require which two parts of the procedure? Enzyme digestion Electrophoretic separation of DNA fragments What are the basic steps of the procedure? Labeled oligonucleotide probes of 15-19 base pairs are added. DNA is amplified in the region of the known mutations, denatured and applied to areas of membrane or filter. What is a Reverse Dot Blot? Variation of Dot Blot procedure How is the Reverse Dot Blot different? The ASO probes are bound to a filter and denatured DNA from the patient is added to the immobilized ASO. Hybridization occurs when base sequences are 100% complementary to the probe What is an example of a southern blot? This procedure is used for analysis of the proximal product of gene expression, mRNA. Cloned DNA probes can determine whether a given gene is expressed and, of so, how vigorously. Northern Blot Blotting Protocol This is a tissue-based molecular diagnostic assay. Common in situ hybridization (ISH), including fluorescent ISH (FISH), probes are used in the diagnosis of hematological malignancies in clouding CMLA, AML, Burkitt lymphoma and other lymphomas. In Situ Hybridization This gene expression profiling uses _____ analysis. A surface (in this example, a glass microscope slide) contains thousands of spots. Each spot contains a large number of DNA fragments. For each spot, the DNA fragments are derived from one Microarrays
Hematology%20PPT%20Flashcards%20Unit%201.docx