Analyzing Protein Part 2 Flashcards
What is the significance of SNPs?
Single Nucleotide Polymorphism explains that we are 99.9% similar to the person next to us but 1 in 1000 nucleotide differences between human genomes.
They can be neutral, predisposing or pathogenic.
Restriction Endonuclease
Cut DNA at specific sites
Don’t need to add SDS in Agarose Gel Electrophoresis since____.
DNA is charged
Ligase Reaction
Gluing DNA together
Genes can be cloned using what ?
Bacteria
Cleave bacteria with restriction nuclease, insert DNA fragment and covalent linkage to create recombinant DNA.
Manipulated gene products can be stored and replicated in ____
Cloning vectors
What is the difference between Genomic Library and cCDNA library?
Genome has everything (exons and introns)
cCDNA (just exons) but requires reverse transcriptase
Both are indefinite storage for material
Why would we use PCR?
Amplify a DNA regions, especially for a sample that is not readily available.
Diagnose HIV, bacterial/fungal infections and gene mutations like cystic fibrosis, thalassemia, hereditary hemochromatosis, Huntington’s and Fragile X
In order to run a PCR, you must know _____ so you can add ____ appropriately for amplification.
DNA sequence
Primers
Detail the steps for PCR
Obtain small sample from patient Denature Add primers Anneal Add dNTPs Taq Polymerase completes replication in a thermocycler
Quantitative PCR or real time PCR is different in that it uses
a probe to fluorescence and quantify a copy number of specific gene.
During denaturing, a complementary oligo with fluorescence tag is used unlike primers
Quantitative PCR or real time PCR benefits
Detect levels of infectious agent
Determine levels of gene expression
What is restriction fragment length polymorphism?
Forensic analysis of DNA evidence and Diagnostics (sickle cell)
All evidence is cleaved with same restriction endonucleases
DNA placed on agarose gel
Begin the Souther blot process entailing:
Add DNA to Nylon Membrane
Add Probe (measures cut size)
Wash off excess probe
A person with Huntington’s disease has several ___ repeats compared to a normal individual.
CAG
DNA Microarrays are used to
determine overall change in gene expression between 2 samples
“dot chip”
