Biology 3594A Lecture Notes - Lecture 23: Auxotrophy, Histidine, Ames Test
13 May 2018
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Department
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Professor
Question 1
Tutorial to show the level of detail expected on each question, similar to questions that are on the final
Some questions will require diagrams, well-labeled diagram is part of the rubric
Don’t have to show what a CNV is but do have to show what is going on
Things you are expected to show, rubric for this question
Expected to draw a flow chart of how CGH looks
Artistic talent not relevant
Labels always help
CGH: show that there is test and control DNA and those are two diff fluorescent labels (here two colours)
Know that you get a mark for another set of control DNA that has a metaphase chromosome spread
Denature and hybridization step for both
Measure relative fluorescent levels of green and red, whether there is a CNV in the segments of DNA
Written portion, talking about benefits and drawbacks
Written by TA
Substantial amount of stuff, be thorough
Make sure you answer the actual question, read the question carefully and get all the info from it to give the right answer
Benefits of CGH: compare methods that are similar (ex. CGH and aCGH, assays that are testing the same thing)
Question 2
We talked about 3 types of ELISAs in class, know the logic behind all 3 and how they originated are
Know the benefits of direct, indirect, and capture/sandwich assay
ELISAs primarily used to measure 8 oxo DG
Large rubrics! Be able to cover a lot of things
Should be able to draw this whole thing out
Say that it’s there, primary antibody, linked to enzyme that catalyzes a substrate to colour reagent
Indirect assay –have secondary antibodies, more enzyme per antigen meaning you can convert more substrate per antigen
That is a good thing because you can amplify the signal, important in terms of sensitivity (good word to know make sure you use it)
Indirect assay biggest advantage: more sensitivity
Souper Soup Question
Ames test for mutagenicity
A picture isn’t good enough for this question, need also a substantial written portion
These are main points, we need to convert into full sentences
Model –say what model is used, specific strain and say that it is a bacterial test
Auxotrophic mutants –explain how the test works, start off with auxotrophic mutants, explain what auxotroph means
Say that they need histidine to grow
Point is to quantity revertant colonies and what revertant is
Prototrophic –go back to original state, can synthesize their own histidine (histidine independent)
Then explain control and test plates, why you need rat liver enzyme, diff between direct and indirect mutagen
Then diagram: not as many points as the writing
castPCR to KASP Assay
Draw a flowchart for both of them
Worth a ton of marks
Tutorial Exam Review Questions
May 13, 2018
3:12 PM
3594 Page 1
Document Summary
Tutorial to show the level of detail expected on each question, similar to questions that are on the final. Some questions will require diagrams, well-labeled diagram is part of the rubric. Don"t ha(cid:448)e to show what a cnv is (cid:271)ut do ha(cid:448)e to show what is going on. Things you are expected to show, rubric for this question. Expected to draw a flow chart of how cgh looks. Cgh: show that there is test and control dna and those are two diff fluorescent labels (here two colours) Know that you get a mark for another set of control dna that has a metaphase chromosome spread. Measure relative fluorescent levels of green and red, whether there is a cnv in the segments of dna. Make sure you answer the actual question, read the question carefully and get all the info from it to give the right answer. Benefits of cgh: compare methods that are similar (ex.
