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- Gel electrophoresis separates molecules based on size O charge O weight What is the restriction cut site for HaellR ATAT GGCC O AATTAA O CGGGCDuring which step in the transformation protocol does the plasmid DNA move into the cell? Heat Shock Grow in LB media at 37°C Plate on media with Ampicillin O Incubate the cells on ice with CaCl2 in the absence of plasmid. fg f1o 19 %24 & 5 6. 7 8. 4 5 R Y U P F G J 2 K 3] vのOne of the frist steps in isolating plasmid DNA via mini-prep is to pellet the cells after O/N culture and then resuspend them in buffer P1. What's the point of pelleting the cells just to resuspend them again? To concentrate the cells P1 is a buffer, preventing small changes in pH O This step is not absolutely necessary P1 begins the lysis process D Question 2 What would happen if we didn't centrifuge the tube containing lysed cells after adding neutralization buffer and instead just added right to the column? O We could fihish the prep but we would have protein/RNA contaminants at the end O The precipitate is less dense than water so it shouldn't get in the way since it floats on top O The precipitate would obstruct the column and our prep is ruined O Our yield would be reduced, but we'd get something
- Why should plasmid sample be mixed in the inverted position and not vortexed? Will these actions affect the plamid DNA extraction?Biology QuestionLooking at the model data i upload and given the question how could i analyse tboth experimental results (antibiotic selection and agarose gel). Including one clearly labelled image of the agarose gel and with an accompanying legend. And how could i interpret the findings of the results and draw a conclusion i.e. what is in each tube? Also is there any further experiments that I could perform to confirm the identity of the plasmid stocks?
- What is the role of the following in the alkaline plasmid screen? G buffer (Cell Suspension Solution) Denaturing Solution (Cell Lysis Solution) Neutralization SolutionExamine the pGLO plasmid DNA solution with the UV lamp. Note your observations. Using a micropipettor, withdraw 10 ul of plasmid and mix it into the cell suspension of the +PGLO tube by pipetting up and down three times. Close the tube and return it to the rack on ice. Also close the -PGLO tube. Do not add plasmid DNA to the -pGLO tube. Why not?The presence of colonies on the bacterial plate post transformation suggests which of the following? Group of answer choices The colonies contain the transformed plasmid and are not resistant to the antibiotic The colonies contain the transformed plasmid and are resistant to the antibiotic The colonies do not contain the transformed plasmid and are not resistant to the antibiotic The colonies do not contain the transformed plasmid and are resistant to the antibiotic
- 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 youDuring the isolation of TOL plasmid from Pseudomonas putida, it was observed that the concentration of the plasmid DNA per 50 mL of sample was extremely low. Describe a technique you could employ to increase the concentration of the TOL plasmid.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.