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Laboratory Procedures for Blood Typing

Laboratory Procedures for Blood Typing

Human Adaptation: Seeking Homeostasis When Under Stress

This lab is a combination of an online blood typing scenario and several exercises where the data

was previously collected.

Scenario 1:

Transfusion with Online Lab

  • The online option is more interactive. If possible, choose between Options 1-3.
  • For Options 1-3, follow the instructions and record data in the online lab book (otherwise you

cannot move to next step). Before you move onto next step, print or screenshot the data you

entered into the online lab book.

  • For Option 4, use the tables provided in the Data Collection handout.

Options:

  • Option 1: Use Chrome:

and select the lab called “Blood Typing”

o If you cannot open, try this: To enable Flash for the site, click the lock icon at the left

side of the Omnibox (address bar), click the “Flash” box, and then click

“Allow.” Chrome prompts you to reload the page—click “Reload.” Even after you reload

the page, any Flash content won’t be loaded—you have to click it to load it.

  • Option 2: Use Internet Explorer:

and select the lab called “Blood Typing”

  • If you cannot open, follow this tutorial: <

player/kb/install-flash-player-windows.html>

  • Option 3: Use Firefox.

and select the lab called “Blood Typing”

  • If you cannot open, follow this tutorial:

flash-plugin-view-videos-animations-games

  • Option 4. Perform this portion of the lab using this paper version:

.pdf >

Complete the lab exercise, fill out Table 1 and answer the lab questions.

Scenario 2 A:

Paternity Question in My Family

True story. When a close relative had her 3rd child she was concerned. One day after the birth,

she asked me if her husband would believe he was the biological father. I was stunned; she is as

straight an arrow as you will ever meet. I had no doubts about her fidelity to her husband.

Blood Laboratory Procedures 2

What made her concerned? Blood types I laughed quietly and then reassured her all was fine.

Fill out Table 2 based on your understanding of blood genotypes, given the blood types

provided. First record all possible genotypes that each blood type could be Then record the

actual genotypes, based on your genetic analysis.

Scenario 2 B:

Paternity Test in the Lab

You have been asked to determine if any one of three men might be the father of a child

(paternity test). A match does not mean that person is the father, but a mismatch means that

person is not. This is simply the first screening test to rule out those men who can’t be the father.

Further testing will be undertaken if the blood type of a sample indicates you can’t rule out a

sample (but not by you).

  • The samples consist of one from the mother, the child, and three candidates (labeled as

Daddy 1, Daddy 2, and Daddy 3).

  • Perform the tests sequentially; complete and record one sample before you proceed to the

next.

  • You must now determine which, if any, of these men could be the father of the child.

Record the missing data in Table 3: Data for Paternity Test Scenario.

Scenario 3:

Fieldwork Determining the Frequency of Blood Types among the HardToGetTo Islanders

You will be studying the blood type frequencies of a population on the isolated island of

HardToGetTo. Their tradition tells of a man who came to live with them who as member of the

EasyToSee Island. This man was quickly accepted by the islanders. Soon, a great plague swept

across the island, killing most of its inhabitants; the man and his extended family, however,

survived. You suspect that the man from EasyToSee Island carried the disease to his new home.

The current islanders claim to be the descendants of the man from EasyToSee Island and the

original islanders who survived the plague. Their culture and language also connect them with

the people of some Islands to the west of HardToGetTo Island, whose blood group frequencies

already have been studied (they are called the WayOutThere Islanders). These archival data are

presented in Table 4 and Table 5.

Some of your data was collected by an earlier team of the Genographic Project (n =76); you will

collect an additional 24 samples by combining your work with that of your classmates (n = 24).

These data are presented in Table 6.

General Overview of Fieldwork Data

  • Some of the data used in this scenario are archival. These data sets were stored in the

archives of the Genographic Project. This means you retrieve the data from these previous

research efforts for the:

o EasyToSee Islanders (recorded in Table 4)

Blood Laboratory Procedures 3

o WayOutThere Islanders (recorded in Table 5)

  • In Table 5 you pick up data collection where the 1st team ended (archival). So, you will

combine the results to determine total samples collected for the HardToGetTo Islanders.

Overview of Analytical Work

  • You will compare the data for the three island groups using a series of three Chi-square

calculations.

  • You are particularly interested in migration patterns and the effects of natural selection on the

blood type composition of the HardToGetTo Islanders.

o The HardToGetTo people think they are closely related to the EasyToSee Islanders.

o You hypothesize that the HardToGetTo Islanders may more closely related to the

WayOutThere Islanders than to the EasyToSee Islanders.

  • Possible hypotheses to explore in this research:

o Null hypothesis: There is no difference in blood type frequencies between the

HardToGetTo Islanders and the EasyToSee Islanders. [Indigenous view]

o Alternative hypothesis 1: There is no difference in blood type frequencies between the

HardToGetTo Islanders and the WayOutThere Islanders [Researchers’ view]

o Alternative hypothesis 2: There is a difference in blood type frequencies between the

WayOutThere Islanders and the HardToGetTo Islanders [Neither view is correct.]

Name ________________________________________________________Anthropology& 215

Grading: For each, use X if completed, P if partially completed, and 0 if not attempted.

___ Table 1 (4 points) ___ Virtual lab questions (3.5 points)

___ Table 2 (2 points) ___ Scenario 2A question (0.5 points)

___ Table 3 (2.5 points) ___ Scenario 2B questions (1.5 points)

___ Table 6 (2 points) ___ Scenario 3 calculations (4 points)

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Scenario 1:

Transfusion with Online Lab

(4 points)

Click on this link to access the online lab called Blood Typing:

1) Complete all steps and print the lab notes OR record the data in this handout. 2) Complete Table 1 below or the tables provided in the online lab notes. 3) Answer the Virtual Lab Questions below or the same questions in the online lab notes.

Table 1:

Clumping Reactions and Blood Types

Blood Sample

Clumping for Anti-A

Antibody

Clumping for Anti-B

Antibody

Clumping for Anti-D

(Rh) Antibody Blood Type

1

2

3

4

Virtual Lab Questions

(3.5 points)

  1. Which, if any, of the blood samples tested can the patient with Type B+ blood receive?

___________________________________________________________________________

___________________________________________________________________________

___________________________________________________________________________

  1. Explain how you were able to use your knowledge of how different types of blood react with

Anti-A, Anti-B and Anti-Rh antibodies to determine the blood types of the four samples.

___________________________________________________________________________

___________________________________________________________________________

___________________________________________________________________________

  1. If a person has Type A blood, he or she would have antibodies for what blood type?

___________________________________________________________________________

___________________________________________________________________________

___________________________________________________________________________

Blood Laboratory Procedures 2

  1. Why is Type O negative blood known as the universal donor?

___________________________________________________________________________

___________________________________________________________________________

___________________________________________________________________________

  1. If a person has Type O blood, what type(s) of blood would she not be able to receive?

___________________________________________________________________________

___________________________________________________________________________

___________________________________________________________________________

  1. Is knowing the ABO blood type of a potential blood donor enough to determine a suitable

match? Explain your answer.

___________________________________________________________________________

___________________________________________________________________________

___________________________________________________________________________

  1. What might happen if someone with Type A received a transfusion of Type B blood?

___________________________________________________________________________

___________________________________________________________________________

___________________________________________________________________________

Scenario 2:

Paternity Test with Synthetic Blood Kit

Table 2: Family Blood Types and Genotypes (2 points)

Person Blood Type Possible Genotypes Actual Genotype

Mother O+

Father A+

Baby O-

Hint: Remember for the Rhesus blood type there are three genotypes (DD, Dd and dd). DD and

Dd are positive, dd is negative.

Paternity Question — Answer the following questions related to Scenario 2A

So exactly what did I say to my relative to ease her mind? (0.5 points)

______________________________________________________________________________

______________________________________________________________________________

______________________________________________________________________________

Blood Laboratory Procedures 3

Table 3: Data for Paternity Test Scenario (2.5 points)

Mom Child Daddy 1 Daddy 2 Daddy 3

Anti-A YES NO YES NO NO

Anti-B NO NO YES YES NO

Rh YES NO NO YES YES

Blood Type

Paternity Questions

Answer the following questions related to Scenario 2B (1.5 points):

Can Daddy 1 be the father? Yes or No; Why or why not?

______________________________________________________________________________

______________________________________________________________________________

______________________________________________________________________________

Can Daddy 2 be the father? Yes or No; Why or why not?

______________________________________________________________________________

______________________________________________________________________________

______________________________________________________________________________

Can Daddy 3 be the father? Yes or No; Why or why not?

______________________________________________________________________________

______________________________________________________________________________

______________________________________________________________________________

Scenario 3: Analytical Comparisons of Fieldwork Data among Three Island Populations

Table 4: Blood Types of All Samples for EasyToSee Islanders (from archives)

Blood Group Total samples Blood group frequencies

A+ 9 0.09

A- 21 0.21

B+ 5 0.05

B- 5 0.05

AB+ 5 0.05

AB- 5 0.05

O+ 15 0.15

O- 35 0.35

Total Samples: 100 Total = 1

Blood Laboratory Procedures 4

Table 5: Blood Types of All Samples for WayOutThere Islanders (from archives)

Blood Group Total samples Blood group frequencies

A+ 35 0.35

A- 15 0.15

B+ 5 0.05

B- 5 0.05

AB+ 5 0.05

AB- 5 0.05

O+ 21 0.21

O- 9 0.09

Total Samples: 100 Total = 1

Table 6: Frequencies for Blood Types of HardToGetTo Islanders (2 points)

Blood Group 1st Team’s samples Your samples Total samples Blood group frequencies

A+ 10 5

A- 15 3

B+ 5 3

B- 5 4

AB+ 5 3

AB- 5 4

O+ 20 0

O- 11 2

1st Team’s Samples: 76 Your Samples: 24 Total Samples: 100 Total = 1

Fieldwork Calculations — Perform the calculations related to Scenario 3:

  • You are to make comparisons between the 3 island populations, using blood type frequencies

as the basis of your analyses. These calculations can be completed outside of the laboratory.

This part of the laboratory is worth 4 points.

o Interpret the Chi-square value for Question 1 (Table 6). (0.5 points)

o Calculate & interpret the Chi-square values for Question 2 (1 point)

o Calculate & interpret the Chi-square values for Question 3 (1 point)

  • Generate 3 hypotheses (1.5 points)

1) In Table 7 observe the Chi-square statistical analysis that compares the WayOutThere

Islander blood types (Table 5) with those for EasyToSee Islanders (Table 4).

2) Researchers used EasytoSee Islander data (Observed Values) & WayOutThere Islanders

(Expected) (1 point)

Answer (check one) (0.5 point)

Does your data support that deviance in your values is by chance alone (nullo

hypothesis)

Does your data support that more than chance influenced the data? (alternativeo

hypothesis)

Blood Laboratory Procedures 5

Table 7: Chi Square Calculation Comparing EasyToSee Islanders and WayOutThere Islanders

O E O-E (O-E)2 (O-E)2/E

A+ 9 35 -26.00 676.00 19.31

A- 21 15 6.00 36.00 2.40

B+ 5 5 0.00 0.00 0.00

B- 5 5 0.00 0.00 0.00

O+ 5 5 0.00 0.00 0.00

O- 5 5 0.00 0.00 0.00

AB+ 15 21 -6.00 36.00 1.71

AB- 35 9 26.00 676.00 75.11

Chi square = 98.54

3) Conduct a second Chi-square statistical analysis that compares the frequencies of HardToGetTo Islander blood types with the frequencies for EasyToSee Islanders). Use the

EasyToSee Islander frequencies as Observed Values. (1 point)

Provide the following:

  • Show all calculations done by hand OR print the Chi-square analysis using the calculator.
  • Your interpretation (the determination).

4) Conduct a third Chi-square statistical analysis of only the HardToGetTo Islanders with the WayOutThere Islanders. Use the WayOutThere Islander frequencies as Expected Value.

(1 point)

Provide the following:

  • Show all calculations done by hand OR print the Chi-square analysis using the calculator.
  • Your interpretation (the determination).

Special Note: Remember, Chi-square uses whole numbers, so use the “Number samples in your

calculations.

5) Generate three hypotheses that might explain the blood type distribution similarities/differences among these three islander populations. Be sure to think/utilize the

forces of evolution as you create your hypotheses (natural selection, gene flow, genetic

drift, and mutation).

Hypothesis #1 (0.5 points):

______________________________________________________________________________

______________________________________________________________________________

______________________________________________________________________________

Blood Laboratory Procedures 6

Hypothesis #2 (0.5 points):

______________________________________________________________________________

______________________________________________________________________________

______________________________________________________________________________

Hypothesis #3 (0.5 points)

______________________________________________________________________________

______________________________________________________________________________

______________________________________________________________________________

Appendix A: Letter of Acceptance for Position

November 7, 2011

Ms./Mr. Anthropology Student 2000 Tower Street

Everett Community College

Everett, WA 98201

Dear Ms./Mr. Anthropology Student,

It is with the greatest of pleasure I inform you of your acceptance to the Summer Internship Program of the

Genographic Project. As you know, this project is sponsored by the IBM Corporation and National Geographic, along with several private sponsors. The Genographic Project is a multiyear research initiative led by National

Geographic Explorer-in-Residence Dr. Spencer Wells. Dr. Wells and a team of renowned international scientists

are using cutting-edge genetic and computational technologies to analyze historical patterns in DNA from participants around the world to better understand our human genetic roots” (National Geographic Society, 2013,

para. 1).

You have been assigned to the serology team that plans to collect samples from HardToGetTo Island, a remote

village in the country of FarFarAway. In that we noted that you have no experience with blood testing (at least

according to your application), we have arranged for you to gain experience through our local hospital, BigBucks and Prestige Providence Hospital. First, you are asked to go through this training as a serologist (Scenario 1). Once

these trainings are completed, you will be assigned to a team of the Genographic Project. At that time, you will be

given a set of archival data — gathered by other researchers (the EasyToSee Islanders and the WayOutThere

Islanders) — against which you will check the data collected from the HardToGetTo Islanders (Scenario 2).

Please, read the enclosed packet of materials prior to coming to our site. We look forward to mentoring you in this

exciting project and to sending you to HardToGetTo Island this summer and receiving the results of your work.

Sincerely,

Cynthia Clarke Director, Genographic Project Coordinator of Student Labor

Suite Fabulous

New York, New York 10000

Reference

National Geographic Society (2013). About, Retrieved from

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Appendix B: Blood Typing

Although one usually hears about type A, B, O, and AB, as well as positive and negative blood, we each actually have not two, but about 29 blood types. Each different blood type is coded for by genes at different loci on our

chromosomes. A blood type is determined by the kind of antigen present on the surface of our red blood cells.

Blood types are useful for tracking genetic traits within and between populations, investigating their biomedical effects, and learning about inheritance.

Blood Laboratory Preparation 2 Required: Visit and print these links as your Appendix B: & this link:

Appendix C: Diagram of Blood Typing Results

Source: Blood. (n.d.) Retrieved from http://english.sxu.edu/~visick/blood.jpg

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Appendix C: Statistical Measures

There are two types of statistical measures: descriptive and inferential. Descriptive statistics are exactly as they

sound: they use numbers to describe the physical world around us. Inferential statistics looks at previous gathered

data to make a prediction about future values.

The most commonly used descriptive statistics are the mode, median, and mean. Each of these is based on a

concept called the central tendency. The central tendency is the idea that a standard bell shaped distribution of

values exists. If this were true, than the mode, median and mean would be the same value (at least in the ideal).

Mode: The most common value. Median: The value were ½ of the values are below and ½ of the values are above this value.

Mean: Is the average value (add up all values and divide by number of measures taken).

Blood Laboratory Preparation 3 Chi square: Is a descriptive statistic that determines whether the observed value is within the acceptable value of

the expected value.

Below is an example of how to calculate the Χ2 (Chi square) value for the averages of five students’ measurement

of a piece of string.

Student Name Observed (O)1 Expected (E)2 O-E (O-E)2 (O-E)2/E

Elliot 123 mm 120 mm 3 mm 9 9/120 = 0.0750

Mark 124 mm 120 mm 4 mm 16 16/120 = 0.1333

Beverly 121 mm 120 mm 1 mm 1 1/120 = 0.0083

Fayla 120 mm 120 mm 0 mm 0 0/120 = 0.0000

Cynthia 118 mm 120 mm -2 mm 4 4/120 = 0.0333

Sum (Χ2) 0.2499

1 Observed values are the means of each student’s three measurements. 2 Expected value is the mean of your three measurements.

Data supports null hypothesis Support alternative

hypothesis

Deviations are not significant Deviations are significant

df ↓ p-value → 0.99 0.95 0.80 0.50 0.30 0.20 0.10 0.05 0.02 0.01

1 0.00016 0.0039 0.064 0.455 1.074 1.642 2.706 3.841 5.412 6.635

2 0.0201 0.103 0.446 1.386 2.408 3.219 4.605 5.991 7.824 9.210

3 0.115 0.352 1.005 2.366 3.665 4.642 6.251 7.815 9.837 11.341

4 0.297 0.711 1.649 3.357 4.878 5.989 7.779 9.488 11.668 13.277

5 0.554 1.145 2.343 4.351 6.064 7.289 9.236 11.070 13.388 15.086

Okay, now you have a value, you need to determine if the Sum (Χ2) is within the expected range for inter-observer error. Below is a Χ2 table of values. The three columns on the right represent the percentage of times this value

should be seen at 5%, 2% and 1% of the time.

The df stands for degrees of freedom and is always 1 less than the number of observations (The number of

observations was 5). The df in this example is 4. Notice the table value that I have bolded; your value is even smaller than this. This is interpreted as greater than 99% of the time; the error value would be an expected value.

Another way to word this is that the variation in values is within expected range so is not the result of anything else,

such as intra-observer error.

Need a table with more df entries? There are many on the Internet, but here is one:

http://faculty.elgin.edu/dkernler/statistics/ch09/images/chi-square-table.gif

Chi-square still a bit confusing? Try reading this tutorial: The Pennsylvania State University. (2014).

The chi-square table. Retrieved from