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Two Quiz Molecular
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How do you get DNA out of the cell?
Mechanically- Blender
Chemically- enzymes and detergents
What are enzymes that glue fragments of DNAcalled?
DNA ligase
What do blunt ends require to keep them stably glued?
T4 DNA ligase (viral)
How do we separate DNA fragments?
Electrophoresis
Restriction Buffer provides optimal conditions with NaCl, Tris-HCl and Mg2+. What do each of these do?
NaCl- correct ionic strength
Tris-HCl- proper pH
Mg2+ - enzyme co-factor
What is a restriction map?
A diagram that shows the location of RE cut site on pieces of DNA sequence
What is a vector?
It is a DNA vehicle to deliver DNA into a host cell
What are the requirements to work with plasmids?
Detect presence of the vector
Direct selection method for cells
Insert genes into vector
Detect the presence of an inserted gene
What is transformation?
Putting the cloned genes into bacteria
What is the charge of nucleic acids?
negative
What is southern blotting used for?
Locate a particular gene
What is PCR used for?
Clinical diagnosis
Genetic analysis
Genetic engineering
Forensic analysis
What do we need for PCR?
Template
Primer (forward and reverse)
Enzyme
Nucleotides
PCR machine
What are the three steps of PCR?
Denaturation
Annealing
Extension
What do we use degenerate primers to find?
DNA sequence of an organism
What are the 2 major steps required for purifying DNA?
Centrifugation- separation by density
Chemical Extraction
What are the two types of restriction enzymes?
Type I- cuts DNA faraway
Type II- middle of recognition site (blunt and sticky end)
Author
icimmy
ID
66063
Card Set
Two Quiz Molecular
Description
Two Quiz Molecular (Ch 9,16,17)
Updated
2011-02-13T19:39:07Z
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