Results and Discussion Report

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

A amp PBR322 4301 fot B Clear Zones Figure 2 The postgraduate student, Demika, inserted her gene of interest into the plasmid, pBR322, before transformation into the competent host cell using heat shock method. After that she cultured the cells on the Ampicillin agar plate before replica plating the colonies onto another Ampicillin (A) and Tetracycline (B) agar plates shown in Figure 2. (1) Referring to the vector pBR322 in Figure 2, which recognition site was cleaved to insert the gene of interest? Based on the observation above, can you identify which colonies are carrying positive mcombinants of BR322? Explain your selection.

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1) Look at the ideal results. Were your predictions accurate, and how did they compare with your results?   2) You used aseptic technique during this lab. Why is it important to work in a sterile manner when working with bacteria in the lab?   3) Why are the cells incubated at 42°C?

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?

5 μL of plasmid DNA (pUC 19) was added to 50ul of CaCl2 competent E. coli that was then added 250 μL of SOC medium, 100 μL of  this solution was plated onto a TSA + ampicillin plate. Can you please show me how I calculate the total number of successful transformants per mL of competent cells plated.  The total number of colnies counted on plate was 70. Thank you

Part ii. ) please

Kpnl 4. Plasmid Z has a size of 7 kb, and the map shows Kpnl (K) and Pstl (P) cut sites relative to each other. This plasmid was digested with three different restriction enzymes 2000 bp 3500 bp Kpnl (K), Pstl (P) and Bgll (B) either alone or in combination and the samples run on an agarose gel as shown below. Where does Bgll (B) cut this plasmid ? Does the plasmid have one recognition site or two for Bg|l? Describe the Bgll cut site in this plasmid relative to the Kpnl cut Plasmid Z -7 kb Pstl site. How many bases to the left or right of the Kpnl cut site would you observe the Bgll cut site. Explain briefly. 1500 bp Pstl Ladder Kpni Psti K/P Bgl K/B KPB 7000 bp 7000 bp 5600 bp 5500 bp 4900 bp 3500 bp 2000 bp 1500 bp 1500 bp 1500 bp 1400 bp 600 bp %3D

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Submit your restriction fragments pUC19 cut by Ava II and Pvu II. Submit your plasmid map with the restriction cut sites labeled. Show all work.

Please answer this asap. Thanks,   You have discovered a new plasmid RK21 in a unique bacterial community. As a first step towardunderstanding this plasmid, you digest the plasmid with three restriction enzymes: SspI, XhoI andSmaI. You run the digested plasmid DNA on an agarose gel, along with an uncut sample of theRK21 plasmid DNA as a control.Unfortunately you forget to load a DNA ladder, and obtain the following results. Assumecomplete digestion of all samples or all the digests worked completely

Choose two genes from Figure 4.6 and draw a graph to represent the change in transcription over time.

Jm109 cells transformed with ligation and plasmid alone is used. The colonies most like:   white blue light blue No colonies

a , b , c question

You first need to create a plasmid. What are the minimal components necessary to develop this plasmid? In addition to these components, please draw the plasmid. In this illustration, I am looking for the components, the direction of transcription (i.e., the direction the genes will be transcribed), and what should be transcribed last? Also, where would you specifically insert the P. falciparum gene in this plasmid, and why are you checking reading frames in your gene? Finally, please name your plasmid using the correct nomenclature too.  You are excited you have this new plasmid; you want to transfect it into P. marinus and provide it as an oral vaccine to laboratory mice. However, even though your supervisor enjoys your enthusiastic attitude, they do not allow you to do this quite yet because all you have is this plasmid. Why doesn’t your supervisor allow you to use the laboratory mouse for this research regarding animal welfare guidelines? Your answer should include the 3 R’s of…

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Write a short discussion based on the result

PCHEM4321. An agarose gel electrophoresis pattern of the plasmid PSPM4321 digestion (restriction) is shown below. Draw a restriction map of a plasmid with the appropriate restriction sites based on the data given below. Hindlll Hindll BamHI +BamHI Figure 1: 1% agarose gel electrophoresis of pCHEM4321 40 24 16 12 12 8 4 4 + |

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...]

a. What is the function of the SDS in the lysis buffer? the NaOH? b. What does the potassium acetate do in the procedure and why?

Please answer the following using the experiment data below. Provide the formulas or methods used for calculations of each.   Number of colonies on LB/amp/ara plate = Micrograms of DNA spread on the plates = Transformation efficiency =   DNA plasmid concentration: 0.08 μg/μl 250 μl CaCl transformation solution 10 μl pGLO plasmid solution 250 μl LB broth 100 μl cells spread on agar 227 colonies of transformants

1. Please help answer this question and please show all work on how you got the answers. Thank you so much!! :)

Solve 5.

Write TRUE or FALSE. If false, write the word/s that make(s) the statement incorrect. 1.The primosome complex includes primase, helicase and gyrase. 2.In plasmid DNA biotechnology, plasmid inserted with a “correct" DNA sequence is transferred to bacteria for rapid.