QUESTION 3You are provided with plasmid DNA at 200ng/uL, 10X restriction digestion buffer, restriction enzymes (1unit/ul) and water. You want to digest 1.5 ug of plasmid DNA with 1 unit (U) of enzyme in 1Xbuffer in a total volume of 80 ul. Complete the table below:SolutionStock solution Working solution Volume neededrestriction digestion buffer 10X1XDNA200 ng/ul1.5 μεenzymes1 unit/ul1 unitWaterTotal80 μL

SolutionStock solutionWorking solutionVolume neededRestriction digestion buffer 10 X1 X8 lDNA200…

You are provided with plasmid DNA at 200ng/uL, 10X restriction digestion buffer, restriction enzymes (1 unit/μL) and water. You want to digest 1.5 µg of plasmid DNA with 1 unit (U) of enzyme in 1x buffer in a total volume of 80 µL. Complete the table below: Solution restriction digestion buffer 10X DNA enzymes Water Total Stock solution Working solution Volume needed 1X 200 ng/ul 1 unit/μL 1.5 μg 1 unit 80 μL

You are provided with plasmid DNA at 200ng/uL, 10X restriction digestion buffer, restriction enzymes (1 unit/μL) and water. You want to digest 1.5 µg of plasmid DNA with 1 unit (U) of enzyme in 1x buffer in a total volume of 80 µL. Complete the table below: Solution restriction digestion buffer 10X DNA enzymes Water Total Stock solution Working solution Volume needed 1X 200 ng/ul 1 unit/μL 1.5 μg 1 unit 80 μL

Human Anatomy & Physiology (11th Edition)

11th Edition

ISBN:9780134580999

Author:Elaine N. Marieb, Katja N. Hoehn

Publisher:Elaine N. Marieb, Katja N. Hoehn

Chapter1: The Human Body: An Orientation

Section: Chapter Questions

Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...

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Calculate the transformation efficiency of an experiment conducted using 10ul of 0.005 ug/ul plasmid, you plated 100ul out of a total volume of 500ul and the count of transformed colonies was 186.
Which of the following scenarios would ONLY occur if your skipped the digest purification step? UV/VIS spectrophotometric quantification of DNA may be skewed by uncut plasmid. Some plasmid molecules may be cut once, by one enzyme, and re-ligate to themselves. The fragments cleaved by the restriction enzymes on the plasmid and insert can re-ligate to their sites, causing reduced ligation efficiency. Plasmid molecules cut with EcoRI and Xbal can ligate to each other instead of the insert.
How many fragments and how big each fragment should be after digesting the plasmid with BamHI? How many fragments and how big each fragment should be after digesting the plasmid with PvuI? How many fragments and how big each fragment should be after digesting with both enzymes? Show your calculation/explanation.
What type of matrix was used to separate DNA in gel electrophoresis? Agarose 1X TBE Glycerol Polyacrylamide Ο 1X TAE
Use 25ul for calculations.
How much 340bp insert DNA should be added to a ligation in which 50ng of 4.3kb vector will be used? The desired vector: insert molar ratio needs to be 1:3.
Why is this the only correct answer? Why doesn't carbonic anhydrase function the best at pH 7?
You have another circular plasmid. Complete and effective digestion of this plasmid with a restriction enzyme yields three bands: 4kb, 2kb, and 1 kb. In comparing the band intensity on an ethidium bromide-stained gel, you notice that the 4 kb and the 2 kb bands have the exact same brightness. The 1 kb band is exactly one fourth as bright as each of these. (Assume there is uniform staining with ethidium bromide throughout the gel.) How many times did the enzyme cut the plasmid? What is the size of the plasmid? Justify your answers to a and b above using a clearly labeled diagram showing the relative location of the cut-sites on the plasmid.
Kha Vu Danels Include: 8DX : Safehy Jor bromie tnto lab repart! Name Section date sheet MAPPING PRACTICE #1 Below is a restriction map for the plasmid PGEN 101 (total length = 20 Kb). Using this map as a guide, give the number of restriction fraqments along with their associated lengths that would result from digesting PGEN 101 with the restriction enzymes EcoRI, BamHI and a combination of ECORI and BamHI. BamHI 3.2 Kb 1.7 Kb EcoRI BamHI PGEN 101 8.7 Kb 5.5 Kb .9 Kb EcoRI ECORI DIGESTION PERFORMED SIZES OF FRAGMENTS OBTAINED 10.4 kb , 0.9kb, 8.7 Kb EcoRI 3.2 Kb, 16. 8kb BamHI EcoRI + BamHI
How many microliters of pGLO plasmid will you need for a PCR reaction is you need 200 nanograms and the plasmid concentration is 0.05 micrograms/microliter?
After plasmid isolation, you take 0.5 uL of the sample and mix it with 1 uL of TE buffer. The absorbance of this sample at 260 nm is 0.8 and the average extinction coefficient of double-stranded DNA at 260 nm is 0.020 (μg/ml)−1 cm−1. What is the concentration of the undiluted sample? 120 μg/mL 80 μg/mL 40 μg/mL 12 μg/mL
Use the gel to answer the following questions. You will be constructing a map of the plasmid, pDiddy. What is the smallest fragment size that the NcoI/EcoRI double digest produces?