AP_Bio_Lab_9_RE_Analysis (1)
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AP Bio Lab 9 Restriction Enzyme Analysis
Modified Lab Report
I am modifying the requirements for this lab. You DO NOT have to turn in a formal lab report. Please
complete the following lab sheet as an informal lab report.
Pre-Lab Activity
To prepare for the following lab, it is important for you to have a good understanding of restriction enzyme
analysis. Review the relevant information in the content, sidebar, and in your textbook.
Materials (all parts)
●
Laboratory Investigations Notebook
●
Metric ruler
Instructions:
This investigation consists of several activities, some of which have been altered for you to conduct this lab at
home. Make sure you understand both the prepared and modified procedure. Begin by reading over the entire
lab (p. 111-124) very carefully and then create investigative ideas that can be tested by experimentation.
Activity I: Restriction Enzymes
Instructions
Read “Activity I: Restriction Enzymes” on p. 113 in your lab manual.
Analysis Questions
:
1.
What is the sequence of the complementary DNA strand (draw it directly below the strand)?
5’-AAAGTCGCTGGAATTCACTGCATCGAATTCCCGGGGCTATATATGGAATTCGA-3’
3’- TTTCAGCGACCTTAAGTGACGTAGCTTAAGGGCCCCGATATATACCTTAAGCT-5’
2.
Assume you cut this fragment with the restriction enzyme
Eco
RI. The restriction site for
Eco
RI
is 5’-GAATTC-3’, and the enzyme makes a staggered (“sticky end”) cut between G and A on
both strands of the DNA molecule. Based on this information, draw an illustration showing how
the DNA fragment is cut by
Eco
RI and the resulting products.
Activity II: DNA Mapping Using Restriction Enzymes
Instructions
Read “Activity II: DNA Mapping Using Restriction Enzymes” on p. 114-115 in your lab manual.
Analysis Questions
:
Consider your classmates. More than 99% of your DNA is the same as their DNA. The small
difference is attributed to differences in your genetic make up, with each person having a genetic profile
or “fingerprint” as unique as the ridges, arches, loops, and grooves at the ends of his or her fingers.
(Be sure to justify your predictions below.)
1.
Based on this information, can you make a prediction about the products of DNA from different
sources cut with the same restriction enzymes?
If DNA from different sources are cut with the
same restriction enzymes then the resulting RFLP patterns may be different.
2.
Will the RFLP patterns produced by gel electrophoresis produced by DNA mapping be the same
or different if you use just one restriction enzyme?
Even with one restriction enzyme, differences
in DNA sequences can still lead to distinguishable RFLP patterns. However, Multiple restriction
enzymes provide a better view of genetic variation.
3.
Do you have to use many restriction enzymes to find differences between individuals?
Using
multiple restriction enzymes can enhance the ability to detect genetic variations between
individuals however it is not always necessary.
4.
Can you make a prediction about the RFLP patterns of identical twins cut with the same
restriction enzymes?
Identical twins share the same DNA sequence, so their RFLP patterns,
when cut with the same restriction enzymes, would be identical. There would be no differences
in the fragment lengths between them
5.
How about the RFLP patterns of fraternal twins or triplets?
Fraternal twins or triplets do not
share identical DNA sequences, as they develop from separate fertilized eggs. Therefore, their
RFLP patterns, when cut with the same restriction enzymes, would likely be different.
Activity III: Basic Principles of Gel Electrophoresis
Inv. 1 – Gel Electrophoresis
Instructions
Read “Activity III: Basic Principles of Gel Electrophoresis” p. 115-122. To complete this part of the lab,
you will complete a modified version by using a virtual lab from the Genetic Science Learning Center.
The procedures in the virtual lab are the same as the directions provided on pages 116-119 of your
student manual, make sure you review both the lab and manual carefully to ensure thorough
understanding.
Procedure
Complete the following virtual lab activity and answer the corresponding analysis questions:
Gel
Electrophoresis
.
Analysis Questions
1.
What 6 items did you need to prepare your gel?
Powder agarose, buffer, a flask, a gel mold, and
a gel comb.
2.
What is the purpose of the buffer in the electrophoresis box?
The purpose of the buffer is to
allow electrical charges flow through the gel.
3.
What is the purpose of loading the DNA size standard (also known as a ladder)?
The purpose of
loading the DNA size standard is to determine old fragments that provide a reference.
4.
What is the charge of DNA?
DNA is negatively charged.
5.
Why do you need to put wells on the negative side of the electrophoresis box?
Since opposing
sides attract, when the current is turned on the DNA will move along the gel towards the
positive.
6.
When you start electrophoresis, what do you look for to start to appear in the buffer that give
you proof that the current is running?
You should look for bubbles to form in the buffer. These
signal that the current is flowing.
7.
What is the purpose of using ethidium bromide or other similar solutions?
Using solutions like
ethidium bromide helps visualize the DNA fragments.
8.
In the picture below
draw
in your DNA sample bands and give your
estimate
for bp size using
the standard as a guide:
Inv 2 – Calculating Size of Restriction Fragments
Instructions
To complete your analysis, read the directions beginning on page 120 of your student lab manual.
Examine the ideal digest below (also on page 121). Using the ideal gel shown in Figure 5, measure the
distance (in cm) that each garment migrated from the origin (the well). (
Hint
: For consistency,
measure from the front end of each well to the front edge of each band, i.e., the edge farthest from the
well.) Enter the measured distances into Table 1. (See * and ** notes below the table for an
explanation for why there are only six bands seen but more fragments.)
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effective dilution produced by the addition of the phage solution?
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Question 3 of 18 -
Bio_U08_USA_FY21
Question: 1-3
A certain plant tissue, typically found in the stem and in the hard outer covering of seeds, contains very thick, rigid cells. Which of the following correctly identifies this tissue and its function?
meristem tissue; gamete formation
O vascular tissue; production of sugar
ground tissue; transportation of materials
O dermal tissue; protection against damage
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39. Ticarcillin 700 mg is ordered. Ticarcillin is supplied as a powder in a vial. Directions
on the vial state “add 2 mL of sterile water for injection. Each 2.6 mL of solution will
contain 1 gram of ticarcillin. How many mL should be adminis
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Basic Laboratory Equipment
Uses/Functions
Picture
1. Microscope
2. Colony Counter
3. Autoclave
4. Microbiology incubator
5. Drying oven
6. Refrigerator (microbiology)
7. Bunsen burner/alcohol lamp
8. Candle jar
9. Anaerobic jar
10. Microhood or Bacteriologic hood/Safety hood/Safety cabinet
11. Bacteriologic filters (Seitz, Chamberlain, Berkfield)
12. Petri dish
13. Culture tubes
14. Hanging drop slide
15. Durham’s tube
16. Staining rack
17. Thermostatically controlled water bath
18. Inoculating loop
19. Inoculating needle
20. Vials
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40. 250 mg Vantin oral suspension is ordered. Vantin is supplied as a powder in a screw-
top bottle. Directions on the label state "add 29 mL water and shake to mix
thoroughly. Once reconstituted, each 5 mL will contain 100 mg of Vantin." How
many mL should be administered per dose?
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electrophoresis)
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(ACADEMIC)
Time
The solid matrix that is usually used to attach the target DNA in southern blotting technique is usually a
O a.
Polyacrylamide gel
O b. Petri dish
Oc.
Cell culture plate
O d. Membrane
Oe.
Micro-titer plate
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14 ▾ BIU A A-
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What are the important characteristics of atoms and elements?
What are the different types of chemical bonds?
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Basic Laboratory Equipment
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Picture
1. Microscope
2. Colony Counter
3. Autoclave
4. Microbiology incubator
5. Drying oven
6. Refrigerator (microbiology)
7. Bunsen burner/alcohol lamp
8. Candle jar
9. Anaerobic jar
10. Microhood or Bacteriologic hood/Safety hood/Safety cabinet
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4. What is the use of ethidium bromide and why must you wear gloves when you handle it?
5. What makes the DNA fragment move towards the positive plate?
6. What is the purpose of glycerol in the sample buffer?
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mead.
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Smear & Stain Preparation
Name: Leidiawa MontaNo
Date:
1. Why are thick or dense smears less likely to provide a good smear preparation for
microscopic evaluation? Please explain.
Becapse, it will diminish the amount ds latcan Pass through by mam
1t difficult to See under the micioscol e, s less liket to Prowde a go
image.
2. What could potentially happen if you leave the slide exposed for too long to the open
flame? Why do you have to be careful?
we can form ring Patterns if we expose the slide for ta0
the flame
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- (ACADEMIC) Time The solid matrix that is usually used to attach the target DNA in southern blotting technique is usually a O a. Polyacrylamide gel O b. Petri dish Oc. Cell culture plate O d. Membrane Oe. Micro-titer plate CLEAR MY CHOICEarrow_forward+ courses/_518299_1/cl/outline?customClassicLocation=%2Fwebapps%2FBb-McGraw Hill-BB5744b9beb8ccb%2Fapp%2Flink%2Finbou.... File Edit Format Tools Help Qa 11 Chapter 2 ● Exam 2 Review Sheet ● Normal ● ● ● Unsaved Chantor 7 Edits will not be automatically saved. Calibri T Save now 14 ▾ BIU A A- EEEE VE What are the important characteristics of atoms and elements? What are the different types of chemical bonds? What are the various types of chemical reactions and solutions? What determines acidity and alkalinity, and how do they relate to the pH scale? What makes carbon the fundamental element of life? What are the four types of biological macromolecules, and what are their general structures and functions?arrow_forwardGive the uses/functions and images of each apparatuses. Basic Laboratory Equipment Uses/Functions Picture 1. Microscope 2. Colony Counter 3. Autoclave 4. Microbiology incubator 5. Drying oven 6. Refrigerator (microbiology) 7. Bunsen burner/alcohol lamp 8. Candle jar 9. Anaerobic jar 10. Microhood or Bacteriologic hood/Safety hood/Safety cabinetarrow_forward
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