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8 Cards in this Set

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Endonucleases usually cleave where?
5’ phosphate/3’ hydroxyl
What is restriction?
Method for cells to differentiate self from non-self

Restriction is the result of an endonuclease and a methylase that each act only upon specific DNA sequences, marking DNA as "self" (okay) and "foreign" (dangerous invader - in the next slide phage DNA)
The vast majority of known restriction enzymes are of what type?
type II


Most type II enzymes cut palindromic DNA sequences
Discovery of restriction enzymes led to the development of ___________ Technology
Recombinant DNA
What is Restriction fragment length polymorphism useful for?
Since each organism (or person) has a unique chromosome sequence the distance between restriction sites varies between individuals: so the length of the fragments varies, leading to polymorphisms.

This can be used to genetically tell individuals apart.
Southern blots and Microarrays are Methods used to do what?
identify DNA fragments that contain specific sequences
What is a DNA Microarray?
solid support onto which the sequences from thousands of different genes are immobilized at fixed locations.

Can be used to:
Compare the genomes of two different species
Compare gene expression levels under different conditions

Sequences are aligned on the support in a specific order.
What are 4 Methods for Determining Protein Concentration?
Determining protein concentration using UV requires that the sample be homogeneous
i.e. only contain a single protein
The Bradford Assay
Dye binding assay
Absorbance shift in Coomassie Blue G-250 when bound to arginine and aromatic residues
The Lowery Assay
Requires two different reactions
The first is a Biuret reaction which reduces Cu+2 to Cu+1
The second uses Cu+1 to reduce the Folin-Ciocalteu reagent (phosphomolybdate and phosphotungstate). Causing a color change. Impacted by tyrosine and tryptophan residues.
The BCA assay
Like Lowery
First a Biuret reaction to reduce Cu+2
Second BCA forms a complex with Cu+1 which it purple colored and is detectable at 562 nm