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QATAR UNIVERSITY

Experimental Biochemistry (CHEM 352)

Report#4

Isolation of Plasmid DNA & DNA Extraction from the beans

Student Name: _____________________                ID: __________________

Lab Partner’s Name: _____________________       ID: __________________  

Date: _____________                                              Section: __________________

Instructor’s Name: __________________________ 


                Part I   Why is the plasmid DNA isolation method you used today called as Alkaline Lysis procedure?            
   Using the plasmid DNA isolation procedure, you used today, complete the following Table regarding the solutions used, their composition and function:  
 Solution #CompositionFunction 
       
         
             
             
  After extracting and precipitating the plasmid DNA, why was it essential to add RNAse to some of the samples?        
 In general, what are the three main features of a plasmid or a vector?   1-   2-   3-       Which is the plasmid you used today?   _________________________  
On the map of the used plasmid Mark (write), the location of the 3 main features that should be present in a plasmid.   As per the plasmid map, which antibiotic resistance marker is present in this plasmid?       On the map, in the MCS (multiple cloning site area) site that is shown, mark and identify (circle around it) the base-pair location of the restriction enzyme used in today’s experiment.  

Part II

  1. Were you successful in extracting the DNA from green peas?  How did you conclude this?
  • From your observation of the isolated DNA as seen in the test tube and, describe the appearance of the DNA you extracted (color, shape, etc)
  •         In the extraction of DNA from green peas, how did the addition of the following reagents help?
  • Sodium chloride:
  • Detergent:
  • Meat tenderizer:
  • Alcohol:
  • Compared to the brand of detergent that was given to you, can other detergents or even soaps be used? If yes, why?  If not, why not?
  • Your group was given 30 g of green peas and the procedure indicated certain quantities of all the reagents.  Can you use more of the green peas to start the experiment?  If so, how would you modify your procedure?
  • Can the same method of isolation of the DNA be used for other raw vegetables, fruits, or other parts of a plant such as leaves? Explain your answer.
  •  Will cooked food, canned, and sterilized food, also be useful for extracting the DNA?