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1. The final elution of your miniprep plasmid DNA has a concentration of 250 ng/uL. You need to load 500 ng on an agarose gel to be able to see the DNA. You plan to assemble a single restriction digest reaction (cutting the plasmid with EcoRI only) with a total volume of 20 uL, and incubate it in a 37oC waterbath for 5 min. What is the minimum amount (in uL) of FastDigest EcoRI enzyme you need to use?
What is the volume of plasmid DNA, in uL, that you would use in the reaction?
2. For Lab 5 he 203L preproom needed to restriction digest a large quantity of lambda DNA in order to have enough for all the students. The total amount of DNA required for one reaction was 22 ug. Starting with the basic protocol, scale up the reaction for the larger amount of DNA. How mouch enzyme (in uL) should be used?
What is the recommended total reaction volume?
3. An SDSU grad student requested a plasmid fom a research lab at UCSF. It expresses a protein of interest from a human gene approximately 242 amino accids in length that has been cloned into a plasmid expression vector called pET15b, sold by Novagen. She decides to do a restriction digest on the plasmid to verify it contains the gene by cutting th vector on each side of where the gene was inserted. The UCSF alb told her the restriction enzymes they used for cloning were Hind III and Ndel. The SDSU lab has the Hind III enzyme in the freezer and doesn't have Ndel. The grad student decides that as long as she cuts it with Hind III and knows the whole size of the plasmid, she can still get data the same day to know if the gene is there. She goes to the internet and downloads a vector map of pET15b to find its size. When she does the digest and runs it on an agarose gel, what size band (bp) should she expect to see if the plasmid has the gene?
For a particular assay, you need cells at a concentration of 300 cells per ml. In your flask of cells growing you counted a sample using a hemacytometer.
Compound with amino groupConsider two small, identical, organic molecules in the cytoplasm of a cell, one with a hydroxyl group (-OH) and the other with an amino group (-NH2).
What is the yield of ATP when each of the following substrates is completely oxidized to CO2 by a mammalian cell homogenate.
Critically discuss the symptoms of a patient needing conscious sedation and how the decision would be made.
A somatic cell of a given plant species contains threebillion base pairs of nuclear DNA and 12 functional centromeres when observed at the conclusion of stage G2 of the cell cycle.
In a germination experiment the germination rate and timing of wild radish (Raphanus raphanistrum) and crop radish (Raphanus sativus) were measured.
Human population doesn't follow a j-shaped curve since humans are able to consciously limit the number of offspring that they produce, which is what usually happens after a population industrializes.
Suppose that the lengths of the double-helical regions and the lengths of the single-stranded regions in them vary within relatively tight limits.
You repeat Snyder's experiment, but instead of determining the ability of each family member to taste PTC, you determine what alleles they carry: 2 PAV (PAV/PAV), 2 AVI (AVI/AVI),
Discuss and explain the differences between aerobic carbon cycle and anaerobic carbon cycle. You can copy the graph from the book.
Assuming that the distance between the corneal surface and the retina is 5 cm in an ostrich and that the refractive power of the ostrich's lens is 5 diopters.
Compute the number of "net ATP" molecules obtained the anaerobic conversion of each of the following sugars to lactate.
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