BglII6. You have a circular plasmid available, with a single RE site for BglII (A^GATCT). You cleave theplasmid with BglII, and join it to both ends of a linear fragment of viral DNA which was obtained bydigesting the viral genome with the RE DpnII (^GATC), resulting in a larger plasmid.☑DpnIITextDpnIIOIf the resulting larger plasmid is incubated with BglII, will the enzyme be able to excise the fragmentof viral DNA? If no, why not? If yes, explain under which conditions. Motivate your answer by showingthe sequence of the plasmid DNA (in black) and the viral DNA (in blue) at both the join positions.Write out the sequences in dsDNA format, if necessary indicate any flanking nucleotides by N. 7.Consider the following plasmid (size 6700 bp), with restriction sites at the positions indicated:BamHI+16700 bp28003500EcoRIBamHIProbe

Approach to solving the question:Steps in answering number 6:Understand the Enzymes and their…

Answered: BglII 6. You have a circular plasmid…

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

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

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A cloning vector map is shown below. EcoRI Bam Ban Hind P-galactosidase Amp Bam Bam EcoRI Ori C Which restriction site is best for inserting a DNA fragment for selection of chimeric plasmid containing colonies? 1) They're all equally good. 2) Hindll 3) EcoRI 4) BamHI
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
1) Given the image below, describe what is happening at each of the 3 sections (i.e. 1, 2, and 3) . [In section 1, the gene donor is providing..., In section 2..., In section 3...]
27. Given the following plasmid map, list how many fragments would exist and their bp length when treated with Hindll and Nde1 Dail 91 Ben BI 51 EstAPI 179 BemBI 2683 Ndel 183 Kasl - Narl - Sfol 235 Bgll 245 Fspl 256 Pul 276 EcoO1091 2674 Aatll - Zral 2617 BiM 2542 Sspl 2501 Prull 306 Benrl 364 Acll 2297 BæAl 387 Apol - EeoRI 396 Xmnl 2294 lacza haall - Sacl - Ecos3KI 402 Accbs1 - Kpnl 400 Awal- BsoN - Smal- TapM I- Ymal 412 Beal 2215 MCS Scal 2177 haml 417 Xhal 0 Pvul 2066 Accl - Hinl - Sall 29 pl. Alau a sb 434 Avall 2059 PUC19 2,686 bp BerDI 1985 Sphi 441 Hindi 447 Acll 1924. Fspl 1919 Prull 628 Avall 1837 Til 641 EsaXI 659 me AllI 1822 Rgll 1813 Bpal 1784 BSPOI - Sapl 683 BsrFl 1779 Bsal 1766 Thl 781 AII - Peil 806 BerDI 1753 Dal 908 Bmrl 1744 ori Aldl 1694 BkiM 1015 BseYl 1110 AlwNI 1217 BeeN 1292 dy
9. Here is a restriction map for a bacterial plasmid showing the cleavage sites for two different restriction enzymes EcoRI and Bam H1. EcoR1 700 bp EcoR1 2000 bp 1200 bp BamH1 Match the banding pattern in each lane of the agarose gel below with one of the treatments listed. Note: Not all treatments listed are run on the gel. Plasmid treatments: A. BamHI+EcoR1 (no RNAse) B. BamHI only (no RNAse) Gel lanes: 1. DNA Ladder = Mol. Wt. Standard (sizes indicated) bp 5000 4000 3000 2000 1650 1000 850 650 500 C. BamH1 + EcoR1 + RNAse D. RNAse only 2. E. BamH1 + RNAse F. EcoR1+ RNAse 3. G. EcoR1 only (no RNase) 4. 1 2 3 4 5 6 5. 6. -
1)Which plate did you see purple/pink/blue bacterial cells? Why did you see this growth? Explain your answer in terms of transformation and plasmids? 2) Calculating Transformation Efficiency For the +DNA/+Amp/+IPTG plate, record the following: Number of transformants (colonies): _________________ Nanograms of plasmid DNA added: 50 ng Final recovery volume: 0.50 mL Volume plated: 0.25 mL Transformation efficiency equation: Transformation efficiency = Number of transformants / µg of DNA x Final volume at recovery (mL)/ volume plated (mL) 3) Using the equation above, calculate the transformation efficiency. 4) Describe the success of the transformation efficiency of this demo based on the calculation you did above?
Examine the structure of the pBR322 plasmid depicted below. Assume total size of the plasmid is 4,361 bp and the blue numbers indicate locations of restriction sites relative to the O point at the top of the plasmid. What size fragments would be generated by the following restriction digestion reactions? 1. Sall 2. Sal 1 + BamH1 3. Sal 1 + EcoR1 4. Sal I + BamH1 + EcoR1 PstI 3607 3000 4000 amp ori HindIII Edit View Insert Format Tools Table EcoRI EcoRV 4359 0 29 185 pBR322 4361 bp 2295 NdeI tet 2000 BamHI 375 651 SalI 1000 Type a short answer in the space provided below.
2005) Bsal 6. On the left is a diagram of the pUC19 plasmid, which is composed of 2,686 bp. Numbers on the plasmid and next to each restriction enzyme cut site represent a base pair (bp) measurement from an arbitrary starting point (0) going in the clockwise direction. On the right is a diagram of an electrophoresis gel. A standard molecular weight (MW) ladder is in the far-left lane, labeling the position of DNA fragments of known sizes from 0.5-10 kb. Lanes 1-5 are DNA bands observed from digestion of the plasmid with one or more of the restriction enzymes. 2500 11500 ori PUC19 2686 bp Sspl (1266) 9 B -HindIII (632) BamHI (662) EcoRI (683) kb 10.0, 8.0 6.0. 5.0 4.0 3.0 2.0 1.5 1.0 <0.5 MW 1 | || 2 3 4 5 a. Which lane(s) shows the DNA fragmentation pattern produced by digestion of the plasmid with either EcoRI, Bsal, OR Ssp1? Briefly explain your answer. b. Cutting this plasmid with restriction enzymes Bsal AND Ssp1 would result in DNA fragments of what sizes? Which lane(s) of the gel…
Competent E. coli cells were transformed with the pGLO plasmid. These transformed cells were then allowed to grow on two different plates: 1) a plate containing LB/AMP and 2) another plate containing LB/AMP/ARA. In which plate would you observe both phenotypic and genotypic changes? Briefly justify your answer. Edit View Insert Format Tools Table 12pt v Paragraph v BIUA e T?v 田 D2
83. A plasmid is cut with the restriction enzyme BamH1 giving fragments of 3000 and 1000 bp as identifiedby gel electrophoresis and ethidium bromide staining. In a seperate restriction digest the enzyme EcoR1gives fragments of 1000 and 1500 bp that are apparent on the agarose gel. What is the most likely size ofthe plasmid in bp? Explain why it's 4000
Construct a restriction map for Plasmid X to show the recognition sites for two commonly used restriction enzymes: EcoRI and BamHI. Plasmid X undigested Plasmid X digested with EcoRI Plasmid X digested with BamHI Plasmid X digested with EcoRI and BamHI 2200 bp 1000 bp 1200 bp 400 bp 1800 bp 200 bp 400 bp 600 bp 1000 bp
5. Consider the following plasmid (size 8000 bp), with restriction sites at the positions indicated: PstI XhoI 7000 8 kb 2000 XhoI Probe 4200 ECORI This plasmid is digested with the enzymes listed below. Indicate how many fragments will be a) generated in each case, and give the sizes of the fragments. PstI XhoI Combination of PstI + XhoI + EcoRI (triple digest) b) Draw the banding pattern you would expect to observe if each of these digestions is loaded into a separate well of an agarose gel, and the fragments separated by electrophoresis. In the first well you load a DNA marker (M) containing fragments with sizes of 1000 bp, 2000 bp, 4000 bp and 8000 bp. This gel is transferred to a membrane in a Southern blot experiment, and hybridised to a c) radioactively labelled 200 bp probe, which anneals to the plasmid DNA at the position indicated on the diagram above. Draw the autoradiographic profile you would expect to observe for the membrane.

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