Quizzes - PGM. 2007. Quiz 1 – 26 September 2007. Please print, use ink, and avoid teensy-weensy printing. Name any one of 2 methods for getting 500 ug of a human gene of your choice other than growing up millions of flasks of cells. PCR, cloning
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Please print, use ink, and avoid teensy-weensy printing.
Remember to put last name first on the cover and the page on which you are writing your answer. Please use pen and avoid teensy weensy lettering.
A) GACGTC or B) ACGT
1. True or False: The restriction enzyme recognition sequences that appear in a useful multiple cloning site do not appear elsewhere in the plasmid. True
There are two major “pieces” of DNA that you need to prepare in the process of making a recombinant DNA construct. What are they? Vector and Insert
Name the enzyme used for:
Last Name, First Name
There are two major “pieces” of DNA that you need to prepare in the process of making a recombinant DNA construct. What are they?
Name the enzyme used for:
5. True or False: The restriction enzyme recognition sequences that appear in a useful multiple cloning site do not appear elsewhere in the plasmid.
A) What adaptors do you use to give your cDNA sticky ends?
a) Pst I b) Bam HI c) you don’t need adaptors
B) What restriction enzyme do you use to get the insert out of your construct after you have amplified and purified enough of your construct to work with? Bam HI
A) covalent or B) non-covalent bonds
The phosphodiester bond created by ligase is
A) covalent or B) non-covalent
In a blot hybridization protocol, what is the word for the labeled DNA that includes your sequence of interest? Probe
What is the word given to blot hybridization in which RNA has been blotted to the membrane from a gel? Northern
If you had only the figure of the Hind III digest shown at right, and knew nothing about the starting DNA, which map(s) on the chalk board might be accurate?
A and B
A, B, and C It was all three, but you had to be there.Quiz 7 – 10 October 2007
Alpha Beta Gamma
A) labeling by chemical cross-linking
B) labeling by random priming
C) both A and B
B) kinase, (and maybe phosphatase first)
5) Which end of a DNA strand is labeled by terminal deoxynucleotidyl transferase (TdT)? 3’ end
By an enzyme.
Autoradiography or digital capture or phosphorimaging
Autoradiography or phosphorimaging
Direct: the light producing enzyme is covalently linked to the probe nucleic acid chain.
Indirect: The enzyme is not covalently bound to the probe. The light producing enzyme is covalently attached to a binding molecule which is in turn allowed to bind non-covalently to the small molecule covalently bound to the probe.
Sequence found somewhere in YFG.
Also accepted: the larger the inserts, the fewer clones. But this answer really addresses the question: Why would you want to use a vector that can accommodate the largest possible inserts? That answer is not really relevant when comparing a genomic library to a cDNA library, because cDNA clone number is dictated by the prevalence of different RNA types, and insert size is dictated by the length of the mRNA.
T or F. A cDNA library made from the mRNA from one type of cell will be the same as a cDNA library made from any other type of cell. False
Different cell types make different mRNAs – that’s what makes them different.
The equation was the one for determining the number of clones that must be plated out in order to have a “complete” library.
F is determined on the basis of the average size of the fragments you will be using as inserts. Average insert size is numerator. Size of the genome with which you are working is the denominator.
Mostly clear placques, because that means most of your plaques have inserts disrupting the lac Z’ gene. (In the case of plaques, the blue color is generated within the bacteria before they have lysed.)
5. Is the translation start site in an exon or an intron? It is in an exon, because the translation start site must be present in the mRNA, and the mRNA includes only sequence from the exons in the gene.
Many wonderful answers.
1. What is it that serves to knock out YF Gene in the procedure for making knockout mice? The G418 resistance gene.
2. True or False – The TK gene will be present in a knockout mouse made by the procedure described in class.
3. Name any one polymorphic phenotype that can be used as a marker to help locate the region of the genome in which a “disease gene” lies. STRs, VNTRs, SNPs, RFLPs. Be sure you know what these letters stand for and what each really is.
4. True or False – The polymorphism in a marker is usually what causes the disease.
5. What method could you use to show that the gene you have found is actually the gene that, when mutated, causes the disease of interest? Check the last several slides of Monday’s lecture.
(There can be more than one answer. Use no more than 3 sentences.)