Introduction for anatomy assignment

Identification through Immunoassay

(Adapted from the Shoestring Biotechnology Project of NABT)

The case:

On Monday night at approximately 9:30 PM you received a call to assemble a team and go to the crime scene where Mrs. Brown's body was found.

Here is the information the police officer shared with you upon arriving to the location:

The body of Farmer Brown's wife was found hidden under a haystack behind the barn at the couple’s farm, east of Plano.

She had been reported missing by one of the ladies in her sewing circle after she failed to show up with her famous lemon tarts at the meeting the day before. The sewing circle lady thought something must be wrong and was very upset that there was no dessert for the sewing circle!

Mrs. Farmer Brown was found after an extensive search of the area. A team of police, highway patrol and volunteers had been searching the area for several hours when a search and rescue dog found something in the haystack.

After the haystack was removed, the body of Mrs. Farmer Brown was found with a knife in her heart! She had been dead since the day before.

The police officer interviewed Farmer Brown and he suggested that a passing stranger must have killed his wife while he was out on the tractor plowing a field. He thought she had gone to the sewing circle and decided to spend the night in town with friends.

The police officer spotted a blood stains on Mr. Brown’s jeans and when he asked about it Mr. Brown said they are from normal farm work. He claims that the blood is either from the dog or the cat he kicked that morning OR from the rooster he killed for last night's dinner!

Your job is to test the bloody jeans to identify if it is human or animal blood and if needed to test a blood sample extracted from the jeans to see if it is the same blood type as Mrs Brown

As a Crime Scene Investigator, you must know the following:

Upon returning to your lab you find that your lab staff (the indispensable Ms. Holmes) prepared the materials you need for the tests (don’t forget to thank her and maybe buy her favorite tea…):

2% agar in a Petri dish

1 soda straw for cutting wells in the agar

Pipettes

1 set of sera (antigen) and antiserum (antibody)

1 permanent marker

Bottles of undiluted yellow and blue dye

You start following the procedure for the immunoassay as follows:

Step 1. Place the Petri plate upside down and divide it into three sections, label them A,

B, and C (see diagram at the end).

Step 2. Label well sites (places you are going to cut holes to fill) like the diagram below.

Measure ~1 cm between the holes in each section.

Prepare the gel:

Step 3. Turn the plate right side up and remove the cover.

Step 4. Fold a soda straw over and squeeze the air out of it.

Step 5. Cut wells in the agar where you marked them (like in the diagram) by sticking the open end of the straw into it and twisting.

Step 6. Release the folded top of the straw and remove the piece of gel (it sucks it up)

Step 7. Repeat until all wells are made in the gel.

Step 8: Load samples (see diagram below)

Put just enough liquid in each well to almost fill it

Key for wells:

Y-yellow

B - blue

S - sample

Po - positive reaction

Ne - negative reaction

Hu - human antiserum (antibody)

D - dog antiserum (antibody)

R- rooster antiserum (antibody)

C - cat antiserum (antibody)

? - unknown from the jeans

Po

Ne

S

D

Hu

?

B

Y

C

R

Step 9. Place the plates aside for 30- 45 minutes.

While waiting, discuss with your team the case and the possible outcome of the immunoassay (see report below). Discuss the different section (A,B, C) on the plate and what each is being used for.

Step 10 - If your team decides to check the blood type – please do so now and add your finding to the report. Please note – Mrs. Brown blood type was reported to be O-

Step 11. – documenting evidence

Take pictures of your results – both the agar plates and the blood typing. Remember, this is evidence and you need to document it.

Take pictures of BEFORE and AFTER and mark the time passed between the two.

Data collection -10/24/2018; Lab report due – 11/14/2018

This is a group project – you should work with your lab partners to complete all parts of the assignment.

· The grade will be a combination of:

· Scientific writing - 80% of the grade

· Peer-evaluation – 20% of the grade

Scientific assignment:

· You will submit one assignment per group

· Assignment should be submitted typed in font size 12, line space 1.5 AND uploaded to the Turnitin.com via canvas (one copy per group). Hard copy of the lab report should be submitted in class on the due date – 11/14/2018

Peer evaluation:

· The form is found on Canvas linked to the assignment

· EACH member of the group should evaluate ALL group members INCLUDING SELF

· The electronic version of the evaluation should be uploaded to Canvas and NO hard copy is needed.

Detailed instructions for the final report:

Introduction:

This Immunoassay uses serum-antiserum tests. (antigen-antibody). You will need to review it and write short paragraph explaining the method.

· You need to explain the principles of the method USING YOUR OWN WORDS. No citations will be accepted

· Keep your description short and to the point – explain the principles of the method without describing your results, without relating the description to your results or without relating to the specific experiment

· Introduction should not be more than a half a page.

Here are some links that can help you with the discussion (those are NOT peer-reviewed and cannot be cited or listed. They are aimed to help you understand the method used):