Frage 1
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A radioactive probe can take what forms?
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ssDNA
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RNA
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Denatured dsDNA
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Oligonucleotides
Frage 2
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Radioactive
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Fluorescent
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Use Ig's
Frage 3
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Name the two ways probes can be designed
Frage 4
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What is codon bias?
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Where cells prefer one codon of an aa to another
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Where cells will reject certain codons due to what they contain
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Where some cells can only make tRNA's for certain codons
Frage 5
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80% of Phe codons are TTC, 20% are TTT
Frage 6
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If the protein is an enzyme it can be found by adding the substrate
Frage 7
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State the possible forms of Ig's for protein separation?
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Immobalised on a column
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Removable from solution
Frage 8
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Probes needn't be 100% accurate to work
Frage 9
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For a Western Blot, how is the DNA stuck to the membrane?
Frage 10
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Once the DNA has been bound, what happens next?
Frage 11
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In an expression vector, where is the new gene added?
Frage 12
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Name the parts on the vector
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Origin
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Antibiotic Resistance
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Promoter
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Terminator
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Multiple Cloning Site
Frage 13
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More compliated vectors have proteins tagged for purification
Frage 14
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What is involved in His Chain Purification?
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The addition of a His chain to the N terminus
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The removal of a His chain from the protein
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The use of a His chain to cut unwatned proteins
Frage 15
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His Purification uses Histadine's affinity for what metal?
Frage 16
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His-tagged proteins will bind to Nickel in a column. Others that don't bind will be washed off.
Frage 17
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The PET system involves using a promoter for the protein that you can control the expression of
Frage 18
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What promoter is used in the PET system to control protein expression?
Frage 19
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In the PET system, what promoter is used to control the T_ expression?
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Lac promoter
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Slo Promoter
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Pap promoter
Frage 20
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What is required for a good shuttle vector?
Frage 21
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Shuttle vectors are usually plasmids with additional sequences
Frage 22
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Shuttle vectors for yeast/fungi need
Frage 23
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A baclovirus would be used as a vector into what?
Frage 24
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Secretion of certain proteins can be caused by what?
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Disulphide bonds
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Acetylation
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Methylation
Frage 25
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Why are there two origins in the Yeast shuttle vector?
Frage 26
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What is the method of selection in yeast?
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URA3- allows growth in the absence of Uracil
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TRA3- allows growth in the absence of thymidine
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GRA3- allows growth in the absence of guanine
Frage 27
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Like bacteria, yeast cannot take up linear DNA
Frage 28
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What does a YAC contain that a plasmid does not?
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Non-useful genes
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Yeast centromere
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Yeast telomeres
Frage 29
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Yeast can allow DNA to be directly inserted into its genome
Frage 30
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How can yeast accept DNA into its genome?
Frage 31
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In order to directly insert a gene, you must flank the gene with identical stretches of the part of the genome you want to insert it in
Frage 32
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Plants would use an aglobacterium T1 plasmid
Frage 33
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How do you replicate transformed plant cells?
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Grown them as plantlets
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Clone them
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Asexually reproduce them
Frage 34
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Mammal shuttle vectors are the same as yeast vectors, but the yeast origin is replaced with what?
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An SV40 promoter
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A UG35 promoter
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A FT12 promoter
Frage 35
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Retroviruses can also be used to insert transgenes into mammilian genomes by packing the transgene in a retroviral capsule