Help
Options
Didn't know it?
click below
click below
Knew it?
click below
click below
Don't Know
Remaining cards (0)
Know
retry
shuffle
restart
0:00
Embed Code - If you would like this activity on your web page, copy the script below and paste it into your web page.
Normal Size Small Size show me how
Normal Size Small Size show me how
MMBIO Quiz 5
| Term | Definition |
|---|---|
| RNA is hard to work with | and unstable |
| we convert RNA back into _____ to make it easier to work with | DNA |
| gene | DNA sequence that encodes a function through RNA or protein |
| gene expression | DNA makes RNA which makes protein which has a function |
| reverse transcription | converting RNA into complementary DNA - cDNA uses reverse-transcription enzyme |
| cDNA | complementary DNA anti-parallel relative to the RNA used as a template to create it |
| reverse transcription steps | mRNA --> primer --> reverse transcriptase --> makes two strands --> alkaline digestion of RNA --> cDNA |
| 3 ways to prime reverse transcription | oligodT/polydT random hexamers gene-specific primers |
| poly dT/oligo dT | TTTTTTTT to make cDNA from all mature mRNA |
| random hexamers | random 6-nucleotide primers to make DNA from all RNAs (primers will bind to everything) |
| gene-specific primers | specific to a certain gene to make cDNA of a specific sequence |
| RT-PCR | reverse transcription PCR of the cDNA (requires specific primers) can all be done in a singular test tube |
| why make cDNA? | DNA is very large due to introns, hard to work with can study alternative splicing using RT you can readily clone mRNA after splicing |
| why clone cDNA | you can make recombinant protein for analysis use to make therapeutic proteins/hormones like insulin |
| q-RT-PCR | reverse transcription PCR TaqMan probing |
| microarrays | used to measure expression at level of RNA can measure expression of thousands of genes at a time |
| microarrays steps | reverse transcriptase to get cDNA microarry chip (thousands of probes in different tiny holes) apply cDNA to chip, hybridize to probes wash away unbound material fluorescence scanner to see how much of each gene was expressed |
| competitive binding | two cDNAs compete for one spot, stronger one will be expressed |
| upregulated | expressed more, brighter on the microarray gene is ON |
| downregulated | expressed less, dimmer on the microarray gene if OFF |
| RNA sequencing | looks at all the sequences within the entire genome |
| RNA sequencing steps | turn RNA into cDNA cut cDNA, attach sequence adapters sequence machine to read sequences compare to reference genome to see where they came from quantify how many came from each gene (up/down regulated) |
| polymorphism | tiny difference in our DNA sequences specific to each person that we can use for identification use: diagnose diseases, identify person |
| human genome is | diploid (2 copies of each chromosome) |
| allele | same genes, different chromosomes because of one from mom and one from dad |
| SNPs | single nucelotide polymorphisms one single deletion, insertion or substitution |
| larger deletions/insertions | yep |
| VNTRs (variable number of tandem repeats) | short tandem repeats, every person has a different number |
| substitution mutant | one nucleotide has replaced another, total is still the same |
| insertion mutant | new nucleotides now present |
| silent mutations | chnage in DNA sequence that does not change aa sequence |
| wobble hypothesis | multiple codons code for the same aa |
| frameshift mutation | insertions or deletions in coding regions not multiple of threes |
| protein coding genes are only a ____ portion of the human genome | small |
| RFLP/forensics | blood --> DNA extraction --> restriction enzymes cleavage --> gel --> blot --> probing --> washed --> X ray use to determine who the culprit is, compare DNA |
| SNPs can have many palindromes or lack common ones | extra cuts - smaller fragments larger cuts - larger fragments |
| level 1 | naked DNA rare 2 nm |
| centromere | DNA sequence repeats where spindle fibers attach |
| telomere | DNA sequence repeats on ends, protects end of chromosomes |
| level 2 | DNA+histone proteins = nucleosome |
| DNA is (-) and histones are (+) | so they attract |
| nucleosome | DNA wrapped around histone proteins |
| ____ - ____ twist of DNA per nucleosome | 1.6 - 2 |
| ___ histones per nucleosome | 8 |
| linker regions | areas between DNA and histone proteins |
| micrococcal nucleases | cuts DNA in linker regions cause it's exposed |
| level 3 | interactions between histone proteins H1 protein greater compaction |
| level 4 | 30 nm fiber further compaction histone H1 protein and accessory proteins |
| acetylation | acetyl group on lysine neautralizes charge, making DNA less attratced to the less positive force loosens DNA |
| acetylated DNA | is loosened and so gene is ON |
| deacetylated histones | deactivated genes - gene is OFF |
| methylation | more compact tighter DNA methyl group blocks acetyl group |
| histone cores | compact DNA |
| histone tails | regulate gene expression |
| HATs | add acetyl groups |
| HDACs | remove acetyl groups |
| ChIP assay procedure | protein and DNA cross link with formaldehyde (glue together) sonication - sonic waves to cut up DNA into smaller fragments immunopreciptation - antibodies pull target down reverse cross link and isolate DNA PCR analysis |
| immunoprecipitation | grab specific proteins with antibodies antibodies are heavy and fall to bottom centrifuge antibody proteins in pellet others light and float away |
Created by:
anyasalmon