Term
|
Definition
| Direct determination of the order, or sequence, of nucleotides is the most definitive molecular method to identify genetic mutations |
|
|
Term
| what are two types of DNA sequencing? |
|
Definition
Maxam-Gilbert Sequencing and
Sanger Sequencing |
|
|
Term
| What is the Maxam-Gilbert Sequencing method? |
|
Definition
| template is divided into 4 tubes, and each is treated with a different chemical, the reducing agent breaks single stranded DNA at specific nucleotides, contents are dried and resuspended in formide (keeps DNA denatured), contents separated on polyacrylamide gel by size |
|
|
Term
| What do the smallest bands on M-G sequencing gel represent? |
|
Definition
| base closest to the labeled end of the fragment |
|
|
Term
| What are bands in the purine and pyrimidine called? |
|
Definition
| whether they are also present in the G- or C- lanes |
|
|
Term
| How is A or T indicated in M-G? |
|
Definition
| by the bands that appear in the G + A lane of C + T lane, but not in the G lane or the C lane |
|
|
Term
| How is G indicated in M-G? |
|
Definition
| present in the G + A lane and the G Lane |
|
|
Term
| How is C indicated in M-G? |
|
Definition
| C + T lane and the C lane |
|
|
Term
| What is the sanger method? |
|
Definition
| primer attached to a P-labeled nucleotide or fluorescent dye nucleotide for detection, ddNTP are added and stops DNA synthesis, the templates and primer are placed in four reaction tubes and dNTPs and one of four ddNTPs are added, DNA polyermase is added, sets of fragments are loaded into polyacrylamide gel |
|
|
Term
| What does the sanger method require? |
|
Definition
|
|
Term
| What is another name for sanger method? |
|
Definition
|
|
Term
| What is special about ddNTP? |
|
Definition
| lack a hydroxyl group found on the 3' ribosome C of the deoxynucleotides |
|
|
Term
| When does DNA synthesis stop? |
|
Definition
| upon incorporation of a ddNTP because without a hydroxyl group at a 3' sugar carbon, the 5-3 phosphodiester bond cannot be established |
|
|
Term
| What does sequencing buffer contain? |
|
Definition
| ingredients necessary for polymerase activity |
|
|
Term
| What does stop buffer do? |
|
Definition
| ends the enzyme activity and denatures the products of the synthesis reaction |
|
|
Term
| what does the smallest (fastest migrating) fragment represent? |
|
Definition
| first nucelotide attached to the primer by the polymerase |
|
|
Term
| What is a newer technology to do the same test? |
|
Definition
| capillary electrophoresis |
|
|
Term
|
Definition
The chemiluminescent signal produced by the action of the enzyme on dioxetane develops in the dark.
Light released by phosphorylation of dioxetane takes place where the probe is bound and darkens the light-sensitive film |
|
|
Term
|
Definition
| Normal inherited suppressor genes require 2 sequential mutations to initiate tumors |
|
|
Term
| What occurs in a germline with an already inherited mutation? |
|
Definition
| a single somatic mutation within a tumor suppressor gene will inactivate both alleles |
|
|
Term
|
Definition
| gene is on chromosome 17, transcription regulator, mutations cause uncontrolled cell growth |
|
|
Term
|
Definition
| A rare inherited disease with a high mortality rate, 50% of affected famlies have extended phenotypes with numerous cancers, characterized by multiple caners in family members, a sarcoma patient and at least 2 relatives with other cancer before the age of 45, only 1 wild type p53 allele |
|
|
Term
| What kind of inhereitance is Li-Fraumeni Syndrome? |
|
Definition
| dominant, genetic predisposition leading to cancer, early onset with multiple primary tumors |
|
|
Term
| autosomal-dominant transmission |
|
Definition
| a child of an affected individual and unaffected mate has a 50%-100% recurrence risk |
|
|
Term
|
Definition
| largest category of disorders and the recurrence risk is 25% |
|
|
Term
|
Definition
| X-linked and are almost always recessive. |
|
|
Term
| women with x-linked disorders? |
|
Definition
| one X chromosome is inactivated in females, the inactivation is reversible so the second copy of X-linked genes is available |
|
|
Term
| males with x-linked disorders? |
|
Definition
| they have only one X chromosome and will likely manifest the disease phenotype |
|
|
Term
| Which labs perform this test? |
|
Definition
|
|
Term
| Which gels are better for sequencing? |
|
Definition
|
|
Term
| what can mutations affect? |
|
Definition
| structural proteins, cell-surface receptor proteins, growth regulators, and enzymes |
|
|
Term
| what do most phenotypes result from |
|
Definition
| multiple genetic and environmental factors that occur throughout life |
|
|
Term
| what do pedigrees determine |
|
Definition
| inheritance pattern of a phenotype through its family members |
|
|
Term
| Three types of DNA mutations? |
|
Definition
| genome, chromosomal, and gene |
|
|
Term
|
Definition
| changes in the number of chromosomes |
|
|
Term
|
Definition
| affect the structures of entire chromosomes which involve movement of large chromosomal regions within the same chromosome or to another chromosome |
|
|
Term
|
Definition
| affect single genes and are often small changes in the DNA sequence |
|
|
