2008_06_05_12_23_08

2008_06_05_12_23_08 - Bib; ID?- Tmfixciaf'fom 5 UIUU'J H....

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Unformatted text preview: Bib; ID?- Tmfixciaf'fom 5 UIUU'J H. Lab #14 7L 1. What is the purpose of restriction digest analysis? «9 6116614 {VJMLQ fll'wmci hmvétx/Lf 79% ,n/g,]gc% 76M . Cl ,/l ': 6 L V1 {fr-glint. r“ 3. What type of gel will be used in today's lab? “vi “(V/0ft”, 3H? 1 4. D - 's POSITIVELY EGATIVEL charged and will run towards 4m; EGATIVE electrode during electrophoresis. 5. What is the function of restriction enzymes in bacteria? 5M0} m éutc‘lflez/TCL fifej/m fir by Group #: '1 Lab 16 1. The 6~histidine segment incorporated before the amin fcarboxy terminus of the protein. 2. Why is the addition of the 6-I1istidinc tag useful? : t? lat-Ml se' «a m l f is m #3 lu gut/ma 3. What metal has an affinity for the 6-histidine tag? 4. To limit non-specific binding. 1' -.M;<l log- is added to both the lysis and wash buffers. (I-Iint: It is also used for clution). 3. The absorbancc Spectrum for your individual fluorosccni protuins will be muzsurLd usmg a {/HC f3? plat/311*; r=%.7_;j,_,_. % but? Half (v. 5., f LA, a alkulfi‘kall‘f’ H {grim/4117 and “dint/57. Group 1-? 1— A Quiz for Lab #15 SW ah r. . 1. What are transformed bacteria? K” "1 “57" If!» t (11A. Hon-l- L1.th 1’1 103 “1 UV”: 54/4 ' . Cells that are treated so that. they are temporarily able to take up DNA are called: -Tftflf‘For‘M-€fli’ Lellf 3. Transcription of the inserted DNA in JM109 bacteria is catalyzed by: W 7? Fa/XM-V‘xtpex 4. The enzyme that catalyzes transcription binds to the 7“], {ammo-JD?” on the pRSETB plasmid. _ A 5. Translation begins at the {hr-I- gor an mRNA and ends at the Hm (£7ng of an mRNA. /" /Extra Credit (1 pt) sinCe my lab will say this is a hard quiz: Define translation in less than 15 words. [rt/fo [447L7vg‘ [I c [ ,‘n 0x. f chfirf Wed re welt/61' Mam? Ma" Ar £4,341. / if. _/ l. A file-f mt‘CJ is a circular double stranded DNA molecule capable of autonomous replication within a hos bacterium. Quiz 9 BIBC [03 2. If our plasmid vector, pRSET B, lacked PT? (bacteriophage T7 promoter} sequence, which of the following molecules would you NOT expect to see? mRNA of our protein W DNA of our protein c. Ribosomes d. T? You diluted your sample l:2000 before reading the absorbence. What is the concentration of your plasmid DNA if your A260 was 0.002? (A260=l.0 for 50ug/ml double stranded DNA). SHOW YOUR CALCULATIONS! ‘ O l . r -_—__.-— w Lu) f lIWl/ _l -- . [7“ 5.000 M, 4. Which ofthe following absorbs UV light with a maximum at 260nm? .00 L f .3 W.” L. X Jo P! :1. Protein Q] Nucleotides (DNAIRNA) c. Ethanol (1. Water 5. Zwitterionic detergents have (POSITIVEINEGATW net charge. a A f g _ Quizfi H < Nat BIBC103 Mma fat, - 1. A enzyme that phosphorylates a substrate. 2. _-E\Efit@lnactivc MAPK (phosphorylated form) helps to maintain the arrest of the un ertilized eggs. (Circle one) . .. (st/(“him HIW/p/fo-e . 3. A2318? IS a llpld soluble molecule that moves across membranes accordlng to concentration gradient. (Be Specific) 4. You mix Sul of 2X protease and phospatase inhibitor cocktail (PPIC) with 20ul of 2X SDS sample buffer. Which of the following information is NOT needed to calculate the final concentration of l’PIC‘? a. volume of protease and phospatase inhibitor c0cktail b. concentration of protease and phospatase inhibitor cocktail 0. volume of SDS sample buffer concentration of SDS sample buffer 5. Where should you be when working with formaldehyde? fume hood b.) on your bench top c. near the sink d. near your TA’s desk Quiz}' 4 Nam: BIBC103 . . . mien!” wet/4e 1. For Western Blottlng, the proteins are separated according to on a polyacrylamide gel using SDS—PAGE. 2. One of the factors that can influence protein transfer to membrane during Western Blotting is the pH of the bufi'er. How does it influence transfer? (state what would happen in alkaline pH versus neutral pH) Em \.(é{4.{f"/\£ LIOWLTAHIH M7r"e- N/t/ft‘eé" (412,44: /4’-~=')4,,4/ “J;an Revival f7 {/“PLKJ “we” f”??? a" film: K 7% lye. xve.n£,/+-'Li “jig-75.37;“ 6Q cflfl/akeffl 3. A protein coated with SDS has a net (POSITVEE GATIVE charge and will run '/ I” toward when voltage is applied. ote: ode=positive terminal, fl; 3% has, . Cathode=nega ive terminal 4 flag . After the protein transfer, what is the purpose of incubating your membrane in blocking baffer? Tb "6/11 “(r-"pv- 145% in to ‘4 m 1% M A6 5. How do you make 800ml of IX buffer with 10X stock solution? [OX {/t 2: (fa-S757 [ox—m) L470 {fjlc’gvzl 3/ (51k; ' . " Lt '1 c/ :GPO [ IN“? W J’V'QML [ M # Utrka ...
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2008_06_05_12_23_08 - Bib; ID?- Tmfixciaf'fom 5 UIUU'J H....

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