Introduction to Biotechnology MethodsStudent Name
Date
Instructions:
1. Please read all of the introduction and background information within the
investigative manual.
a. Once you have done so answer the prelab question BEFORE
completing any of the lab’s activities.
2. Once you have completed the prelab questions proceed to the activities
of the lab within the investigative manual.
a. As you read through the instructions for completing each activity
make sure you also:
i. Complete any instructions (append photos, etc)/ and answer
any questions found in the post lab questions for each
activity.
ii. Take the photos of your experiments in each activity as
directed below. IMPORTANT: Don’t clean-up your lab until
you know what portion of the experiment you need to take a
picture of.
3. Here is a video that will introduce you to the lab and its main concepts.
The student is encouraged to watch it.
a. Introduction to Biotechnology
Prelab Questions
1. PCR, as described in the investigative manual, is a very important and
commonly used procedure in biotechnology and biochemistry labs. In
2020 a new virus, SarsCoV2, caused a world-wide pandemic. One of the
diagnostic tools used to detect if a person was infected by this virus was
(and is) PCR. Describe how PCR can be used to detect the presence or
absence of SarsCoV2. Is this method 100% reliable?
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2. In table 1 of the investigative manual, you are given five restriction
endonucleases, their recognition sites, and cleavage pattern. Shown
below is a DNA sequence before exposure to Cla1. This sequence is
labeled “No Fragmentation” and has the recognition site of Cla1 in bold
text. Below this is the sequence after exposure to Cla1 showing fragment
1 (in blue) and fragment 2 (in red).
No Fragmentation
5’ – GAGACCATCGATACCGGC – 3‘
3’ – CTCTGGTAGCTATGGCCG – 5’
Fragment 1
5’ – GAGACCAT
3’ – CTCTGGTAGC
Fragment 2
CGATACCGGC – 3‘
TATGGCCG – 5’
Scientist have discovered significantly more restriction endonucleases
than what is shown in your investigative manual. BamH1, for instance, is a
commonly used restriction endonuclease with the recognition and site
and cleavage pattern:
BamH1 Recognition site
5’ – GGATCC – 3‘
BamH1 Cleavage pattern
5’ – G/GATCC – 3‘
5’ – CCTAG/G – 3‘
In consideration of this produce the fragments of the following DNA
sequence if it were exposed to BamHI:
No Fragmentation
5’ – GAGACCGGATCCACCGGC – 3‘
3’ – CTCTGGCCTAGGTGGCCG – 5’
Fragment 1
2
Fragment 2
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3. Earlier in the semester we ran a paper chromatography experiment on
the pigments commonly found in chloroplast of plants. In this lab you are
being exposed to gel electrophoresis. In the space below state the
similarities and differences between these two lab experiments.
4. Before proceeding to activity 1, please read the preparation, activity 1,
and activity 2 instructions in the investigative manual. Having done so
proceed to activity 2 and fill out the purpose and hypothesis section
before beginning activity 2.
Activity 1
Instructions:
1. Open the investigative manual. Locate all the needed materials
supplied in the kit and those you will need to supply yourself.
2. Lay them out in your work area.
3. Read through the entire set of instructions found in the investigative
manual for the activity to avoid making mistakes when you go to
execute the experiment.
4. Once you have read through the instructions go back to step 1 and
begin executing the experiment.
5. Please answer the questions below and/or append appropriate
representations of data (photos, graphs, etc). REMEMBER don’t clean
up until you have taken the appropriate photos of your experiment as
described below.
Photo 1 – Activity 1
Take a picture and insert the image(s) of your completed gel diagram from step
5.d. of the “Sickle Cell Anemia Detection Simulation: PCR, Restriction Enzyme
Digest, and Electrophoresis” section in activity 1 of the investigative manual:
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1. DdeI produces sticky ends. For the wild-type beta-globin sequence, how
many DNA fragments are present in the following digestion by Ddel?
2. For the mutant beta-globin sequence, how many DNA fragments are present
in the following digestion by Ddel? If the number of fragments is different
than that of the wild-type beta-globin sequence following Ddel digestion,
explain why.
3. For the Ddel-digested, wild-type beta-globin sequence, draw out the
fragmentation pattern. How many basepairs are in each fragment?
Remember that a base pair includes two bound nucleotides. If a base is not
paired because of a sticky end, do not count it.
4. For the Ddel-digested, mutant beta-globin sequence, how many
nucleotides are in each fragment? How many basepairs are in each
fragment? Remember that a base pair includes two bound nucleotides. If a
base is not paired because of a sticky end, do not count it.
5. On the basis of fragment size, how can the difference between the wildtype sequence and the homozygous mutant sequence be recognized?
6. What fragments would be present following Ddel digestion of a sample from
someone with a heterozygous beta-globin genotype?
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7. On the gel diagram, indicate where the DNA fragment(s) in the Ddeldigested beta-globin samples would be expected to run. Draw a line for
each band. Use the DNA marker with known base pair sizes to orient the
bands. A Ddel-digested sample from a newborn with an unknown betaglobin genotype is drawn in the last lane.
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8. Based on the gel diagram, what is the genotype of the unknown sample
from the newborn? Explain your answer.
Activity 2
Instructions:
1. Open the investigative manual. Locate all the needed materials
supplied in the kit and those you will need to supply yourself.
2. Lay them out in your work area.
3. Read through the entire set of instructions found in the investigative
manual for the activity to avoid making mistakes when you go to
execute the experiment.
4. Once you have read through the instructions go back to step 1 and
begin executing the experiment.
5. Please answer the questions below and/or append appropriate
representations of data (photos, graphs, etc). REMEMBER don’t clean
up until you have taken the appropriate photos of your experiment as
described below.
Purpose statement: (This should be the question the experiment is attempting to
address. It should be written as a question.)
Hypothesis statement: (This should be an “if/then” testable prediction that
addresses the question/purpose of the lab.)
Evidence/Claim statement: (This should be a statement regarding whether your
hypothesis was supported or refuted and what data/evidence allows you to
make this claim.)
Reflection statement: (This should be a statement of what you learned, how your
understanding changed, if you have new questions, and what connections can
you make between the lab and the content in the book and other
assignments.)
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Photo 1 – Activity 2
Take a picture and insert the image(s) of your completed gel from step 14. Of
the “Mini Gel Electrophoresis of Dyes” section in activity 2 of the investigative
manual:
1. Having now completed activity #2 go back to the evidence/claim and
reflection statements and complete them.
2. Draw a diagram of the gel that was loaded with dyes, including which dye
was put in each lane. Draw the bands that appeared in each lane following
electrophoresis. Indicate the color of each. Which combination of dyes
make up each of the two unknown samples?
3. Given the results of your gel, what do you think is the charge of each
molecule in the dyes? Which molecule do you think has the largest
molecular weight, which has the smallest?
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Photosynthesis and Plant Pigments Lab Report
Abstract
Photosynthesis is the interaction by which plants produce natural atoms from carbon
dioxide and water within the sight of light. It permits plants to change over daylight into
synthetic energy, additionally requires these shades. The light energy frees electrons from water
atoms, which join with CO 2 to frame O2 in the light-reliant responses. The electron transport
chain and photosynthesis are two instances of biochemical cycles that utilize photosynthesis.
Chlorophyll is the shade found in green growth and plants that handle photosynthesis. These
interesting little atoms handle the dynamic varieties we find in all that from blossoms to passes
on to products of the soil. Without them, plants would be restricted to shades of green.
It can build the pace of photosynthesis through higher light openness or expanded carbon
dioxide fixations. In this analysis, we are keen on testing how both the light openness and
different carbon dioxide focuses will influence the pace of photosynthesis.
Purpose statement: Will the rate of photosynthesis in leaves increase when it is under various
light conditions or CO2 concentrations?
Hypothesis statement: If the light conditions are higher and the CO2 concentration is lower,
then more photosynthetic activity will occur.
Evidence/Claim statement: My hypothesis was supported. Based on the evidence taken from
the experiment, more disks floated when put into the light that they did in the dark experiment.
Variables:
Independent variable: light and dark environment; dependent variable: leaf disks; and the
controlling variable: time in minutes
Introduction
The course of photosynthesis is significant for the endurance of the two plants and
creatures. Plants convert light into synthetic energy, which they used to make glucose from
carbon dioxide and water. It then involved the glucose for development and cell fix. To an
extreme or too little, carbon dioxide can repress the photosynthesis cycle, prompting lower plant
yields, hindered development, and even passing. Photosynthesis is the interaction that permits
plants to change over light into put away energy, an indispensable advance in the carbon cycle. A
few natural variables, including temperature and stickiness, can influence the pace of
photosynthesis. The effects of carbon dioxide and sun openness on photosynthesis have been the
focal point of this investigation.
Materials and Methods Used
The materials used for this analysis are dish cleanser, regular water, coin, light source,
paper towels, pencil, 5 to 8 plant leaves (spinach), Timer, scissors, little bowl brimming with
water, 2 receptacles, 20ml. gradated chamber 25 ml, plastic test tube, baking soda, medication
cup, straight pin, ruler, test tube rack, oil pen, dropper pipet, and 4 needles.
Strategy/Method
Preparation:
It refrigerated the leaf plate passes on to safeguard the shade of the leaf until I prepare it
for the trial. It ought to disinfect working regions prior to the use of the materials for the
experiment. Set of the capacity region for light by utilizing a work area light or a light with 60
watts.
Then, at that point, gets ready dim stockpiling region. Set up a foamy water using a drop
of dish cleanser into 250ml of water. Place it in a medication cup. Then, at that point, add baking
to the 250-ml container and filled it with refined water. Mix using the dropper pipet. Ensure that
the combination subsequent to blending is suspended. Before it will settle out, measure 5ml of
baking soft drink arrangement in 25 ml graduated cylinder, then empty it into the second
container. Mark it with 0.24%.
Place the 10 leaf circles to the 4 needles by string straw and situating it near the tip of the
needle. Unclogger is supplanted and gradually pushes the plate towards the needle tip. Fill the
needles with the 0.24% arrangement from the container. Place the tip of the needle into the
water, then pack it up to 6 ml mark. I tapped the air pockets out. Then mark the pre-owned 4
needles as A, B, C, and D.
Light Experiment
In this analysis, a holder loaded up with water is ready. I lay 40 to 50 leaf circles on the
hard surface, which is safeguarded with a paper towel. Plastic drinking straw is used to cut the
leaf into a plate. By tenderly crushing and bending the finish of the straw, the circles ought to be
taken out and guarantee that the inside the plates are not harmed. Then, at that point, place it in
the water until the examination will begin.
Result
Under the light, the circle which is in the water (control arrangement) stays at the base.
Notwithstanding, the circles with a treatment arrangement (baking pop) ascent as it uses CO2
and go through photosynthesis, consequently creating oxygen bubbles. The air pockets made the
circle float. In the wake of eliminating in the light and setting the cups in obscurity, the treatment
circle quit going through photosynthesis, and the leaf plate sank.
Discussion
During the dim as well as the light preliminaries, the recognized factors are as per:
Independent variable: light and dim climate; subordinate variable: leaf circles; and the
controlling variable is the time.
The drifting leaf plates gave a visual portrayal of what occurred during the
photosynthesis. The arrangement used made more carbon dioxide, and made the leaf plate sink.
The utilization of baking soda arrangement expands the pace of photosynthesis. If the 0%
of the baking soft drink arrangement will be applied in the trial, there will be no expansion in the
photosynthesis rate. In the event that seriously baking soft drink arrangement will be used, the
leaf plates will surface faster because it expands the carbon dioxide level. This initiates the
photosynthesis under the light.
.Conclusion
I did not realize what a huge different setting of light and air made when it comes to
photosynthesis until this experiment. I realized that baking soda made it hard for the disks to
float because it produces carbon dioxide. Baking soda makes it hard for the disks to float because
it produces carbon dioxide. I could not observe thoroughly a photosynthetic rate without
understanding the process at which it takes the plants to produce oxygen. Having the baking soda
as a comparison to the other mixture makes it easier to understand the process of photosynthesis.
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