genetics midterm

why do we incubate our plasmid with competent bacterial cells on ice for thirty minutes

so the negatively charged plasmits will be attracted to the positively charged bacteria

in the equation C1V1=C2V2 what does C1 stand for

concentration of stock solution

SDS is a ___ that works by taking apart_____

Detergent; phospholipids

how many milliliters of a 50x buffer will you use to make 8L of 1x buffer

160mL

which of the following would be added last to a reaction

Hind III

killing all the bacteria on an agar plate except the specific ones you want is called

selection

if you walk by an electrophoresis gel running at 50V it is a good bet that the researcher is trying to

get a clearly readable gel for a publication

how many bands should have been seen in your lane of gel if our digest with EcoR1 and BamH1 worked properly

two

the purpose of TE buffer in this project is to

elute the DNA

what is the only reason you would raise the temperature of a DNA reaction to 95 degrees C

to break hydrogen bonds

why do we use ethanol in a DNA wash solution

so the DNA will precipate on the spin column membrane

knowing the number of hydrogen bonds between pairs in DNA, which of the following sequences would take the most energy

GGAATTTACCCGTGT

short DNA strands travel____ than longer DNA strands and will therefore be a ____ distance from the well

faster: farther

what is the smallest increment a 200uL micopipettor can transfer

0.1uL

how many grams of agarose would you add to a 150mL of TAE Buffer to make a 2% gel

3g

why is it important to quickly add the neutralization solution after cell lysis solution

to prevent the release of the bacterial chromosome

why do we use SYBR safe in a cloning procedure

Invitrogen SYBR Safe DNA Gel Stain is a highly sensitive dye for visualizing DNA in agarose or acrylamide gels. SYBR Safe stain is specifically formulated to be a less hazardous alternative to ethidium bromide that can be viewed with blue-light or by UV excitation

the GT mismatch in codon 12 of the human ros gene is implicated in about 80% of ____ cases

bladder cancer

explain the steps for making bacteria competent for transformation

you need the bacteria in log phase

treat with CaCl2, this makes the outer positive making the bacteria competent, meaning that it has a much thinner membrane and able to manipulate easier

in order to transform a bacterium, the cell must be

competent

how much SYBR safe dye would you add to a 500mL gel

50uL

why was an ssDNA used as a control in the electrophoresis of our annealed products?

ssDNA, by definition, is single-stranded, so it won't be able to form the double-stranded structures that your annealed products are supposed to be. This allows you to verify that your annealing process is actually working, and that the products you see on the gel are indeed the result of the annealing reaction, and not just the presence of the ssDNA control.

the pSB2 plasmid is 6kb long and contained a kanamycin resistance gene. digesting the plasmid with AatIII and nar I yields a 0.7kb cutout. digesting the plasmid with Nar I and Not I yields a 0.85kb cutout. you are given a protocol to cut 0.77ug of pSB2 with Aat III and Not I and elute into 50uL. what is the final concentration of the gel extracted DNA?

your T4 ligase enzyme just arrived from introgen . it is a 50uL volume containing 250 units of the enzyme. the accompanying literature states that the enzyme must be stored in a storage biffer with the following conditions: 10mM Tris-HCL (pH 7.5), 10mM KCL, 1mM DTT and 50% (v/v) glycerol. you wan tthe enzyme at a final concentration of 1 unit/uL and you have the following stock solutions: 0.1M Tric-HCL, 0.1M KCL, 10mM DTT, 100% glycerol. calculate how much of each reagent you will need to store your enzyme. ( this is a real world application and professor white does this every semester when this enzyme comes in so its ready for you to use.)

• 25 µL of 0.1M Tris-HCl

• 25 µL of 0.1M KCl

• 25 µL of 10mM DTT

• 125 µL of 100% glycerol

• 50 µL of the T4 Ligase enzyme

ligation is:

the joining of two peices of dsDNA end-to-end