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Molecular midterm

QuestionAnswer
Major components of PCR rxn? Template, Taq, primer set, MgCl2, KCl, DNTPs (nucleotides) for DNA synthesis
What components of loading buffer for DNA and protein do? It's for the reducing agent, Tris, Glycerol, Dye and SDS
What DNA intercalator and stains do? Gel Star? Visualization? It is used as fluorescent probes to visualize double and single strands DNA, oligonucleotides and RNA in gels.
What about the structure of FCF green allows it to absorb at 625nm max? Extended conjugation system within the molecule lowering energy requirement
Why should RuBisCo elute in high salt? RuBisCo binds strongly to resin
Dialysis moves molecules through osmosis, not diffusion. False
Using two antibodies for western blotting increases signal. True
All proteins absorb at 215nm strongly; Most proteins absorb at 280nm due to residues (W, Y, F) True
The largest DNA amplicons (bp) travel fastest to the bottom of the gel because of charge False
What does magnesium chloride do in PCR? Cofactor for Taq, Stabilizes primer/template duplex
What does glycerol do in loading buffers? Increases viscosity and weight
EDTA is a monovalent cation chelator that sequesters any sodium or potassium in solution. False
Make a .8% (w/v) agarose gel at 85ml for a product of interest that's 5000bp .68 grams of agarose in 85ml
Artificial Selection produces GMOs. False
PCR has leading and lagging strand synthesis like DNA replication. False
What structure in DNA allows for quantification at 260nm? Nucleosides
CamV 35S is commonly said to be a gene by biologists? False
SDS is an anionic detergent that denatures proteins True
TUB1 (Tubulin) is commonly said to be a gene by biologists? True
The template or copy number for our PCR was 1. False
Why was our pellet resuspended in 100ul TE instead of 5ul when that's all we needed? To do reruns, To dilute contaminants
Anionic exchange columns such as Q-Sepharose are specific only for RuBisCo False
What is the purpose of fully air drying after ethanol precipitation? To prevent ethanol from interacting with the PCR rxn
The Air around you is a solution. True
Which pipette is best used for 235ul? p1000
40 cycles of PCR means? 40 denaturing, annealing, and extension steps
Make 2.5% agarose at 150ml in 1X TBE. 500ml of 10X TBE is provided 3.75 grams agarose in 150ml 1X. 15ml of 10X +135ml diH2O
Make 200ml of 20mM EDTA from .5M EDTA master stock. 5ml of .5M EDTA in 195ml
What is the difference between master stock and working stock? Master stock is high concentration for storage, Working is used in reaction
How much .2M glucose can be made from 500ml of 1M glucose? 2.5L
Do a V/V calculation of ethanol and make 1L 70%. You are given 1L 100% ethanol. 700ml 100% ethanol and 300ml water.
Adding solutes into water you must subtract the grams from the total volume. For 1L 1000ml-700g (sucrose)= 300ml water False
What does the term "amplify" or "propagate" refer to in molecular biology? To increase the number of copies of a specific DNA sequence
For PCR, Taq polymerase does not have proofreading ability but KAPA HiFi, Q5, and Phusion polymerase have proofreading True
What was the template DNA for our GMO+ control? Genomic
For PCR, what is meant by polymerase and chain? Enzyme that synthesizes DNA; one reaction feeds into the next
Primers are short 5bp synthetic single strands pieces of DNA False
What does a biological catalyst do? Speeds up reactions by stabilizing transition states/position substrates
Adding ammonium sulfate too quickly can produce microenvirnments that precipitate RuBisCo out early. True
For anion exchange, what is the charge of the resin/slurry/column? Positive
For making solutions using solid mass, you always "fill to" final volume True
Make 750mL of 0.75M NaCl (3M stock), 0.5M Tris (M.W.: 121g/mol) 187ml NaCl, 45g Tris, Fill to 750ml water
Salting in is the pushing proteins out of solution with a high concentration of salts. False
Salting out is the pushing proteins out of solution with a high concentration of salts. True
Once you have a standard curve equation, it's infinite and you can input any values in science (not math) False
Most molecules that absorb light, only absorb light at a single discrete wavelength. False
Because DNA absorbs at 260nm and proteins (most) absorb at 280nm, they also emit light we can see. False
What component of Edward's buffer allows for lysis? +boiling 10% SDS
Match what property of RuBisCo was used to separate it Salting out; Solubility, Chromatography;Charge, PAGE;Size
Find the dilution factor for: 1/10 serial dilution 7 times. 1*10^7
OD is given in micromolar concentrations. False
Find DNA concentration when OD= .94 47ng/ul, 47ug/ml
Find purity ratio for DNA using OD260/280 for .73 and .36 respectively 2.0 Ratio; Pure
GelStar is an interclator/fluorophore with a planar structure because of the benzene rings True
SDS-PAGE loaded proteins go from anode to cathode and agarose gels pull DNA from cathode to anode False
Bromophenol blue is a tracking dye in many loading buffers that travels at: 500bp Agarose; 10kDa PAGE
Heating TE/RNase A Buffer to 60C would be bad as it denatures the RNase. False
Know how to do DNA purity ratios for only 260/280. Know what’s considered pure? Anything more than 1.8. Though above 3 is sus. Anything below is 1.8 is not pure
Know how to calculate for DNA concentration from a given OD 260 value just multiply with 50 to get answer.
w/v problems. How to make agarose gel 1% or 2%? 1%= 1g/100ml solution
v/v problems. Dilute ethanol from 200 proof (pure) ethanol. 1%= 1ml/100ml.
For the above either know how to setup proportions with whatever percent/100ml = X/final volume. convert percent to decimal moving the decimal twice to the left and multiply with volume.
Created by: zahriabeshea
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