Molec Cell Lab Final

A L-2 micropipette can transfer liquids from 0.1-2.0 microliters. If the L-2 volume indicator reads 015 then how much volume does this indicate?

0.15 microliters

What is the maximum absorbance that the spectrophotometer can reliably read a sample?

1.0

A L-20 micropipette column indicator reads 1-0-0. What volume does this indicate?

10.0 micorliters

What end of the primer do free nucleotides bind?

3' end

Each sample being measured in this video contains the protein Fast Green FCF. What wavelength is the FAST Green FCF protein being measured at?

625 nm

How many times would you need to blank the spectrophotometry if measured the wavelength of a sample 400-700 nm at 50 nm intervals?

7

If 1 mL of a 4-Fold diluted sample is mixed with 1 mL of water, what would be the concentration of the resulting solution?

8-Fold dilution

A p20 pipette should be used to acquire 21 ul of liquid True or False

False

GMO stands for "Genes Mended in Organisms" True or False

False

Molarity is defined as the number of moles of solute that are dissolved in 1 mL of solution True or False

False

Multiple restriction enzymes can recognize and cleave the same restriction site in a DNA sequence True or False

False

Restriction enzymes recognize specific sequences called genes True or False

False

The EDTA in this solution is considered the solvent (rather than the solute) True or False

False

The absorbance of a sample as measured by UV-visible spectrometry is inversely proportional to the concentration of the sample, such that, as the concentration increases, the absorbance decreases. True or false

False

Why os GelStar added to the agarose solution?

GelStar illuminates the DNA in the gel

To make a 50ml of 0.8% agarose solution in 1X TBE we need ___ g of agarose i. need the molar mass of agarose to solve this ii. 1.4 g iii. 0.4 g iiii. 0.9 g

III. 0.4 Gg

Which of the following precipitates DNA?

Isopropanol

Which solution is the DNA in when the sample is ready for PCR?

RNAase Buffer

Looking at the spectrum of the sample shown, what can you say?

The sample contains both DNA and protein

Why was a soil sample added to the PCR mixture?

The soil is being tested for the presence of DNA

If a solution has a pH that is below the desired pH, then:

The solution needs to be titrated with base

What is the purpose of the thermocycler for performing PCR?

The thermocycler adjusts the temperature of the samples according the the prescribed PCR protocol

Why is an electrical current applied to the gel?

To move the DNA which travels towards the positive charged anode

An amplicon is a PCR product, and the DNA to be amplified is the template True or False?

True

An uncut plasmid can exist in different forms, which can result in several bands in the control lane True or False?

True

During ion-exchange chromatography, samples that had the highest protein concentration were saved. These samples were then analyzed further and prepared for SDS-PAGE. True or False

True

In the gel electrophoresis the smaller fragments migrate at a faster rate compared to the larger fragments True or False

True

PCR is a DNA amplification process True or False

True

Restriction enzymes are named according the the bacteria that produce them True or False

True

Serological pipettes are used to transfer liquids exceeding 1 mL True or False

True

To calculate the real concentration of an undiluted solution, you have to multiply the calculated concentration of a diluted solution with the dilution factor. True or False

True

You should NEVER rotate the volume adjuster on a micropipette beyond the upper and lower limit. True or False

True

pH describes how acidic or basic an aqueous solution is True or False

True

When do you press to the second stop on the plunger on a micropipette?

When displeasing the liquid from the micropipette tip

What kind of food is being tested for being a GMO is this experiment?

banana

When do primers bind to the DNA being tested?

during the cooler temperature, usually around 55˚C

You want to make a solution with 2mM Tris in 50 ml total volume. How much tris will you add in 50 ml of dH2O? F.W of Tris= 121.14 g/mol i. 0.0121 g ii. 1.21 g iii. 0.00121 g iiii. 121 g

i. 0.0121 g

To prepare 50 mL 1.5% agarose gel in 1X TBE, how much of each reagent will be needed? i. 0.75 g of agarose, 5 mL 10X TBE, and 45 mL dH2O ii. 0.075 g of agarose, 50 mL dH2O iii. 75 g of agarose, 50 mL of dH2O iiii. 7.5 g of agarose, 5 mL of 10X TBE and 45 mL of dH2O

i. 0.75 g of agarose, 5 mL 10X TBE, and 45 mL dH2O

One milliliter equals to how much μL? i. 1000 ii. 0.1 iii. 100 iiii. 10

i. 1000

What would be the matching strand to the following palindromic sequence: 5'-GGATCC-3' i. 3'-CCTAGG-5' ii. 5'-GGATCC-3' iii. 5'-CCTAGG-3' iiii. 3'-GGATCC-5'

i. 3'-CCTAGG-5'

If a linear piece of DNA has four sites for a particular restriction enzyme, into how many fragments will that restriction enzyme cut the DNA? i. 5 ii. 2 iii. 3 iiii. 4

i. 5

The best tool to measure 146 μl solution is _____ i. P-200 ii. P-2 iii. P-1000 iiii. P-20

i. P-200

The biological role of restriction enzymes is to: i. act against bacteriophages and other foreign DNA ii. restrict the damage to DNA by ultraviolet light iii. limit the size of DNA in certain bacteria iiii. make bacteria resistant to antibiotics

i. act against bacteriophages and other foreign DNA

Restriction Enzymes are isolated from: i. bacteria ii. virus iii. protozoa iiii. fungi

i. bacteria

Strength of interaction between proteins and beads is based on ______ and ______ of charges on the protein. i. number, location ii. none iii. type, size iiii. size, amount

i. number, location

SDS-PAGE isolates proteins based on their: i. size ii. charge iii. solubility iiii. if the proteins can bind to an antibody or not

i. size

When using a micropipette, if you push to the second stop to fill it, you will get _____. i. too much solution ii. too little solution iii. either too much or too little solution iiii. the correct amount of solution

i. too much solution

We need 500mL of a 20g/L solution of NaCl. How much mass of NaCl do we need to add to make this solution? i. 100 g NaCl dissolved in 500 mL of dH2O ii. 10 g NaCl dissolved in 500 mL dH2O iii. 100 g NaCl dissolved in 50 mL dH2O iiii. 1g NaCl dissolved in 500 mL dH2O

ii. 10 g NaCl dissolved in 500 mL dH2O

If a circular piece of DNA has 4 sites for a particular restriction enzyme, into how many fragments will that restriction enzyme cut the DNA? i. 3 ii. 4 iii. 6 iiii. 5

ii. 4

We know that the PCR steps are carries out at different temperatures. At what temperatures does the denaturation step occur? i. 72˚C ii. 94˚C iii. 55˚C iiii. any temperature

ii. 94˚C

What is the name of the substrate that we used during western blotting to induce a color change? i. DNPH ii. BCIP/NBT iii. Sodium azide iiii. DCIP

ii. BCIP/NBT

What micropipette will you use to measure out 140 μl? i. L-1000 ii. L-200 iii. L-20 iiii. L-2

ii. L-200

Which of the following is true regarding the steps of PCR? i. denaturation occurs at 95˚C ii. all are correct iii. annealing is the step where the two primers bind with the target gene sequence complementary iiii. tag polymerase is needed in adding the DNTPs in making the new DNA strand

ii. all are correct

Restriction enzymes are ____ i. catalyze the addition of a phosphate group to certain proteins to activate them ii. are specific ribonucleases, which can degrade mRNA after its synthesis iii. on the cellular membrane and restrict the trafficking of macromolecules into the cell

ii. are specific ribonucleases, which can degrade mRNA after its synthesis

If a protein is large and highly negatively charges it will require a high or low salt concentration to elute it from an ion-exchange column? i. medium salt concentration ii. high salt concentration iii. low salt concentration iiii. NaOH wash solution

ii. high salt concentration

Select the affirmation that is incorrect about pH: i. pH is defined mathematically as pH= -log10 [H+] ii. pH is a measure of the carbon ion concentration in a solution iii. the structure and function of macromolecules is greatly affected by the pH of the solution iiii. the pH of a solution tells whether the solution is acidic, basic, or neutral

ii. pH is a measure of the carbon ion concentration in a solution

What is correct about plasmids? i. serves as the only genetic material for most of the organisms ii. they are extrachromosomal iii. they are fungi, that can cause diseases in humans iiii. they are single stranded

ii. they are extrachromosomal

GMO plants contain additional gene(s) to allow the plants to: i. taste more like meat ii. tolerate adverse farming conditions iii. functions like insects iiii. function like humans

ii. tolerate adverse farming conditions

The slope of a standard curve of Abs vs. [protein] is equivalent to which Lambert-Beer variable? i. OD ii. ε iii. c iiii. D

ii. ε

Which of the following formulas should be used to dilute a solution from high to low concentration? i. C=M/V ii. M=C/V iii. (Ci)(Vi)=(Cf)(Vf) iiii. y=mx+b

iii. (Ci)(Vi)=(Cf)(Vf)

What is the molarity of 5g of NaOH in 750 mL of solution? MW of NaOH is 39.99 g/mol. i. 0.0167 M ii. none iii. 0.167 M iiii. 167 M

iii. 0.167 M

Make a 500 mL solution using 30 g/L KCl i. 100 g KCl ii. not enough information to determine the amount iii. 15 g KCl iiii. 20 g KCl

iii. 15 g KCl

How many copies of target gene strand will you have after 3 cycles? i. 32 ii. 4 iii. 16 iiii. 8

iii. 16

What wavelength does DNA absorb the most light at? i. 200 nm ii. 280 nm iii. 260 nm iiii. 400 nm

iii. 260 nm

What defines the UV spectrum of light? i. < 400 nm ii. > 400 nm iii. 200-400 nm iiii. none of the above

iii. <400 nm

In order to correctly load a micropipette with fluid you must ____ i. push the eject button ii. turn the white wheel on the pump upward iii. add a tip and push the hunger to the first stop iii. add a tip and push the plunger to the second stop

iii. add a tip and push the plunger to the first stop

A DNA has too much protein contamination, the OD260/OD280 ratio of the sample will be ______. i. higher than 1.8 ii. 1.8 iii. lower than 1.8 iiii. lower than 1.8

iii. lower than 1.8

DNA moves from the _______________ end to the ____________________ end in the gel electrophoresis because DNA is ____________________charged. i. positive, negative, negatively ii. postitive, negative, positively iii. negative, positive, negatively iiii. negative, positive, positively

iii. negative, positive, negatively

A useful technique for making millions of copies of a specific region of DNA is: i. gel electrophoresis ii. spectrophotometry iii. polymerase chain reaction (PCR) iiii. restriction enzymes

iii. polymerase chain reaction (PCR)

In nature, the purpose of restriction enzymes is to: i. protect the bacteria from replicating its DNA at the wrong time ii. protect the bacteria from virus attack by not allowing the virus to attach to the cell iii. protect the bacteria from virus attack by cutting up foreign DNA iiii. protect bacteria from the DNA of other organisms that bacteria infects

iii. protect the bacteria from virus attack by cutting up foreign DNA

The purpose of the reaction buffer (Tris-HCl, KCl, and MgCl2)) in the PCR mixture is to: i. it includes the dNTPS ii. provides the DNA template for Taq polymerase to bind to iii. provide a stable environment (pH, ions, cofactors) for Taq polymerase iiii. allow conductivity of DNA to move from the cathode to the anode

iii. provide a stable environment (pH, ions, cofactors) for Taw polyermase

When using a micropipette, if you depress the plunger to the second stop to draw the sample, you will get i. the correct amount of solution ii. too little solution iii. too much solution iiii. either too much or too little solution

iii. too much solution

How many grams of NaOH are contained in 1500 mL of 0.25 M NaOH solution? (NaOH MW=40 g/mol) i. 2.345 g ii. 2.785 g iii. 3.170 g iiii. 0.375 g

iiii. 0.375 g

To make an 80 ml 0.8% agarose gel for DNA electrophoresis, how much agarose do you need? i. 80 g ii. 0.8 g iii. 6.4 g iiii. 0.64 g

iiii. 0.64 g

How many antibodies did we use in the protein isolation experiment? i. 3 ii. 1 iii. 0 iiii. 2

iiii. 2

Determine the amount of ammonium sulfate needed to reach 50% saturation level if you have 32mL. i. 27.2 g ii. 6.72 g iii. 272 g iiii. 2.72 g

iiii. 2.72 g

Plasmid DNA is treated with restriction enzyme A, and three bands are seen on an agarose gel following electrophoresis. This plasmid therefore has ___ restriction sites for enzyme A. i. 4 ii. 0 iii. 2 iiii. 3

iiii. 3

If the recognition sequence of the restriction enzyme HindIII is AAGCTT, then how many covalent bonds will be broken by the enzyme in the following double strand DNA molecule? AAGCTTGAAGCTT i. 1 ii. 2 iii. 3 iiii. 4

iiii. 4

Which would be the best to describe the five components of PCR? i. DNA template, Taq polymerase, Primers, DI water ii. DNA template, DNTPs, RNA polymerase, Primers, Buffer iii. DNA template, Taq polymerase, RNA, Primers, Buffer iiii. DNA template, DNTPs, Taw polymerase, Primers, Buffers

iiii. DNA template, DNTPs, Taw polymerase, Primers, Buffers

What does the color on top of each micropipette plunger refer to? i. which lab group the pipette belongs to ii. types of chemicals that can be transferred by the pipette iii. the colors mean nothing iiii. size of the plastic pipette tip to use

iiii. Size of the plastic pipette tip to use

The OD260/OD280 ratio of a DNA sample is 1.96. The DNA sample is most likely ______. i. contaminated by protein ii. contaminated by RNA iii. cannot be predicted iiii. a pure sample of DNA

iiii. a pure sample of DNA

Restriction Enzymes were primarily used as: i. all are correct ii. defense against protozoa against antibiotics iii. tool by virus to cleave bacterial genome iiii. defense by bacteria against bacteriophages viruses

iiii. defense by bacteria against bacteriophages viruses

What is the purpose of adding sodium dodecyl sulfate (SDS) to your samples? i. washes off the salt ii. prepares the sample to be separated by density iii. regenerates the sample iiii. denatures the protein structures and adds negative charges to the protein

iiii. denatures the protein structures and adds negative charges to the protein

The representation of Beer Lambert's law is given as OD = εcd. If 'd' represents distance, 'c' represents concentration and 'OD' represents absorption, what does 'ε' represent? i. color intensity ii. concentration of the sample iii. path length iiii. extinction coefficient

iiii. extinction coefficient

Positive controls are used to intentionally produce the expected result. Internal controls ensure that the experimental procedures are working as expected. Tubulin is used as a(n) _____________ control. i. external ii. positive iii. negative iiii. internal

iiii. internal

Molarity (M) is measured un what units? i. L/mol ii. g/l iii. mol/mL iiii. mol/L

iiii. mol/L

What is the unit of absorbance which can be derived from the Beer Lambert's Law? i. l*gm-1 cm-1 ii. l*mol-1 cm-1 iii. cm iiii. no unit

iiii. no unit

Generally, agarose gels are used to distinguish DNA samples, while SDS polyacrylamide gels are used to distinguish ________________ of different sizes. i. nucleic acids ii. lipids iii. carbohydrates iiii. proteins

iiii. proteins

What does a spectrophotometer directly measure? i. the amount of a chemical in a material ii. what wavelength of light we are seeing iii. the weight of a material iiii. the amount of light that a substance absorbs

iiii. the amount of light that a substance absorbs

The secondary antibody binds to which two sites? i. the gel and milk ii. the membrane and milk iii. the substrate and primary antibody iiii. the primary antibody and alkaline phosphatase

iiii. the primary antibody and alkaline phosphatase

Which of the following is NOT a dNTP used for PCR? i. dCTP ii. dATP iii. dTTP iiii. dGTP iiiii. dUTP

iiiii. dUTP

Plasmids are: i. small, self-replicating, extrachromosomal, circular, double-stranded DNA ii. often carry genes for antibiotic resistance iii. can not be transferred from one bacterium to another iiii. all of the above iiiii. i and ii only

iiiii. i and ii only

What are some application of restriction enzymes? i. they can be used for DNA finger printing ii. they can be used to assist insertion of genes into plasmid vectors during gene cloning iii. they can be used in mass protein production experiments iiii. all of the above iiiii. i and ii only iiiiii. ii and iii only

iiiiii. ii and iii only

What is an amplicon?

the DNA sequence being targeted

What happens when the PCR cycle reaches 95˚?

the double stranded DNA separates

Which pole do the lanes in the gel need to be closest to?

the positive pole (anode)

In the solution making and Spec module, the TRIS in this example is considered:

the solute

Why is Edward's buffer added?

to break apart membrane structures, releasing the contents of the cell

Why does the agarose need to be microwaved?

to get the agarose to dissolve into the buffer

The Tris-base reagent acts as a buffer. What is the purpose of buffers in solutions?

to stabilize the pH of a solution