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8 Cards in this Set
- Front
- Back
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Endonucleases usually cleave where?
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5’ phosphate/3’ hydroxyl
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What is restriction?
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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) |
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The vast majority of known restriction enzymes are of what type?
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type II
Most type II enzymes cut palindromic DNA sequences |
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Discovery of restriction enzymes led to the development of ___________ Technology
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Recombinant DNA
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What is Restriction fragment length polymorphism useful for?
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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. |
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Southern blots and Microarrays are Methods used to do what?
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identify DNA fragments that contain specific sequences
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What is a DNA Microarray?
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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. |
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What are 4 Methods for Determining Protein Concentration?
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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 |
