HOSA Biotechnology Event Guidelines (August 2023) Page 1 of 17
New for 2023 – 2024
Skill events now require attire appropriate to the occupational area be worn for Round 2.
These guidelines are written for ILC. States may modify events or have different event processes and deadlines.
Be sure to check with your Local/State advisor (or state website) to determine how the event is implemented for the
regional/area or state conference.
Editorial updates have been made.
Event Summary
Biotechnology provides members with the opportunity to gain knowledge and skills required for a laboratory setting
using biotechnology. This competitive event consists of two rounds. Round One is a written, multiple-choice test
and the top-scoring competitors will advance to Round Two for the skills assessment. This event aims to inspire
members to learn more about biotechnology careers.
Sponsorship
This competitive event is sponsored by Bio-Rad Laboratories, Inc.
For resources, helpful videos, and support specifically designed for HOSA and this competitive event, please visit
www.biorad.com/hosa
Dress Code
Competitors shall wear proper business attire or official HOSA uniform or attire appropriate to the occupational area
during testing. Round 2 skill events require attire appropriate to the occupational area be worn. Bonus points will
be awarded for proper dress.
Competitors must provide:
• A photo ID
• Two #2 pencils (not mechanical) with eraser
• Ruler (metric, w mm marks)
• Glasses, safety glasses, face shield or goggles
• Closed-toe shoes
• Disposable non-latex gloves
• Lab Coat (Optional)
HOSA Biotechnology Event Guidelines (August 2023) Page 2 of 17
General Rules
1. Competitors in this event must be active members of HOSA and in good standing.
2. Eligible Divisions: Secondary and Postsecondary/Collegiate divisions are eligible to compete in this
event.
3. Competitors must be familiar with and adhere to the “General Rules and Regulations of the HOSA
Competitive Events Program (GRR)."
A. Per the GRRs and Appendix H, HOSA members may request accommodation in any
competitive event. To learn the definition of an accommodation, please read Appendix H. To
request accommodation for the International Leadership Conference, submit the request form
here by May 15 at midnight EST.
B. To request accommodation for any regional/area or state level conferences, submit the request
form here by your state published deadline. Accommodations must first be done at state in order
to be considered for ILC.
4. All competitors shall report to the site of the event at the time designated for each round of competition.
At ILC, competitor’s photo ID must be presented prior to ALL competition rounds.
Official References
5. All official references, including websites, are used in the development of the written test and skill rating
sheets. In addition, some skills have supporting video resources to help competitors prepare for
competition per item #14 below.
6. Brown, J. Kirk. Biotechnology A Laboratory Skills Course. Bio-Rad. Latest edition.
7. Starr and Taggart. Biology: The Unity and Diversity of Life AP. National Geographic Learning
Cengage. Latest edition.
8. Biotechnology Careers
Round One Test
9. Test Instructions: The written test will consist of 50 multiple choice items in a maximum of 60 minutes.
10. Time Remaining Announcements: There will be NO verbal announcements for time remaining during
ILC testing. All ILC testing will be completed in the Testing Center and competitors are responsible for
monitoring their own time.
11. Written Test Plan
• Biotechnology industry practices and careers .............................................................4%
• Biotechnology in health……………………………………………………………………...4%
• Governmental regulation of biotechnology………………………………………………...4%
• Basic laboratory skills .................................................................................................14%
o PPE
o Preparing solutions (calculations, use of balance and other equipment)
o Pipetting
• Microbiology and cell culture ......................................................................................12%
• DNA structure and analysis.........................................................................................14%
• Bacterial transformation...............................................................................................10%
• Polymerase chain reaction (PCR)................................................................................14%
• Protein structure, function, and analysis .....................................................................14%
• Immunological applications..........................................................................................10%
12. The test score from Round One will be used to qualify the competitor for Round Two.
HOSA Biotechnology Event Guidelines (August 2023) Page 3 of 17
13. At the International Leadership Conference, HOSA will provide basic handheld calculators (no graphing
calculators) for addition, subtraction, division, multiplication, and square root calculations.
14. Sample Round One Test Questions
1. What type of bond connects nitrogenous base pairs and holds the two strands of a DNA
molecule together? (Bio-Rad pg 114)
A. Hydrogen
B. Nitrogenous
C. Oxygen
D. Carbon
2. Which discipline of systems biology investigates the full complement of DNA in a cell? (Bio-Rad
pg 5)
A. Microbiomics
B. Proteomics
C. Genomics
D. Metabolomics
3. What was the first bacterium used commercially to produce genetically engineered
human insulin? (Starr pg 240)
A. Saccharomyces
B. E. coli
C. Epstein-Barr
D. Staphylococci
Round Two Skills
15. Round Two is the performance of a selected skill(s). The Round Two skills approved for this event are:
Textbook (Bio-Rad)
Time
Allocated
Video
Resource(s)
Skill I: Using Micropipets,
Transfer Pipets, and a Balance
pp. 50-53 (Part 3), 381, and 383
(use of balance)
15 min
Videos 1 and 2
Skill II: Restriction Digestion
Reaction
p. 140 (Part 1)
15 min
none
Skill III: DNA Gel Electrophoresis
pp. 140-141 (Part 2) 391
20 min
Video
Skill IV: DNA Gel Interpretation
pp. 136-138, 142, 392
15 min
none
Skill V: Bradford Protein
Quantitation Assay
pp. 254-255 (through step 10), 395
20 min
Video
Skill VI: Bacterial Transformation
pp. 167-171, 392
20 min
Video
Skill VII: Calculation of
Transformation Efficiency
pp. 155-156, 393
10 min
none
Skill VIII: Qualitative ELISA
pp. 314-316, 400
20 min
Video
(FOR ALL SKILLS, ANY BODY FLUIDS WILL BE A SIMULATED PRODUCT)
16. The selected skill(s) will be presented to competitors as a written scenario at the beginning of the
round. The scenario will be the same for each competitor and may include a challenging component that
will require the competitor to apply critical thinking skills. Where appropriate, scenarios will also provide
protocols to those they could expect to see if working in an industry laboratory (these protocols will not be
as detailed as those provided in the textbook; students are expected to know basic skills such as
micropipet use, labeling of tubes, avoiding sample cross-contamination, lab hygiene, and basic workflows).
A specific Biotechnology sample scenario can be found HERE.
17. Timing will begin when the scenario is presented to the competitor and will be stopped at the end of the
HOSA Biotechnology Event Guidelines (August 2023) Page 4 of 17
time allowed.
18. The scenario is a secret topic. Competitors MAY NOT discuss or reveal the secret topic until after the
event has concluded or will face penalties per the GRRs.
19. Judges will provide information to competitors as directed by the rating sheets.
20. Competitors must use all equipment correctly and safety. Judges will stop a student and not award points
for a step if they see a competitor is about to cause a risk of harm or about to damage equipment or other
supplies.
21. Selection of the correct micropipet for a specified volume is an essential skill. Though other micropipets
exist (2, 10, or 100 µl, for example), the skills in this competitive event are to be performed using a 20, 20-
200, or 100-1,000 µl micropipet (a p20, p200, or p1000).
22. The protocols for these skills follow protocols given in the Bio-Rad textbook, Biotechnology A Laboratory
Skills Course (Brown JK). Note that, in skill VIII, the incubation times have been shortened to enable
competitors to complete the skill and see color development within the allotted time.
Final Scoring
23. The competitor must earn a score of 70% or higher on the combined skill(s) of the event to be
recognized as an award winner at the ILC.
24. Final rank is determined by adding the round one test score plus round two skills score. In case of a
tie, the highest test score will be used to determine the rank.
HOSA Biotechnology Event Guidelines (August 2023) Page 5 of 17
Biotechnology
Section # _____________________ Division: ______ SS _____PS/C
Competitor # __________________ Judge’s Signature ______________________
*For all Judge verification steps, full points are only awarded if all components are accurate.
Though the 100-1,000 µl micropipet could be used for these steps, this skill specifies use of a 20-200 µl micropipet (p200).
Skill I: Using Micropipets, Transfer Pipets, and a Balance
(Time: 15 minutes)
Possible
Awarded
1. Entered testing area wearing closed-toed shoes and donned proper PPE:
glasses/safety glasses/goggles, and gloves (lab coat is optional).
2 0
2. Labeled 3 microcentrifuge tubes: for example, p200, p1000, and TP.
4 0
3. Weighed each tube and recorded its mass on the paper provided.
*Judge verified competitor cleared the balance before weighing each tube.
4 0
Using the 20-200 µl micropipet
1
4. Pipetted 200 µl of the liquid provided to the tube labeled for p200 use.
*Judge verified competitor (i) selected 20-200 µl micropipet, (ii) set it to 200 µl, (iii) used an
appropriate pipet tip, and (iv) transferred liquid without air bubbles or losing sample in the
transfer process.
4 0
5. Repeated Step 4.
2 0
6. Pipetted 100 µl of the liquid provided to the tube. Closed the tube tightly.
*Judge verified competitor (i) set the 20-200 µl micropipet to 100 µl, (ii) used a proper pipet
tip, and (iii) transferred liquid without air bubbles or losing sample in the transfer process.
4 0
Using the 100-1,000 µl micropipet
7. Pipetted 500 µl of the liquid provided to the tube labeled for p1000 use. Closed the
tube tightly.
*Judge verified competitor (i) selected 100-1,000 µl micropipet, (ii) set it to 500 µl, (iii) used
a proper pipet tip, and (iv) transferred liquid without air bubbles or losing sample in the
transfer process.
4 0
Using the transfer pipet
8. Transferred 500 µl of the liquid provided to the tube labeled for transfer pipet use.
Closed the tube tightly.
*Judge verified competitor filled the transfer pipet to the appropriate line without air
bubbles and transferred the entire volume to the tube.
4 0
Calculating mass of liquid
9. Weighed all three microcentrifuge tubes and recorded mass of each on the paper
provided.
*Judge verified competitor cleared the balance before weighing each tube.
2 0
10. Calculated the mass of the liquid in each tube (in g) and recorded it on the paper
provided.
*Judge verified calculated mass and that the mass of liquid in each tube was ~0.50g (the
mass of the liquid transferred using the transfer pipet may differ). Judge confirmed correct
units (g) and number of significant figures were used (for example, 0.50 g).
2 0
HOSA Biotechnology Event Guidelines (August 2023) Page 6 of 17
Skill I: Using Micropipets, Transfer Pipets, and a Balance (cont.)
Possible
Awarded
Cleaning up
11. Cleaned work area:
a. Disposed of pipet tips, microcentrifuge tubes, and transfer pipet in
waste receptacle.
2 0
b. Cleaned work area with surface disinfectant.
2 0
c. Removed PPE.
2 0
d. Washed hands or used alcohol-based hand-rub for hand hygiene.
*Judge verified steps 11 a-d were performed in the order written here.
2 0
TOTAL POINTS - SKILL I
70% Mastery for Skill I = 28.0
40
HOSA Biotechnology Event Guidelines (August 2023) Page 7 of 17
Biotechnology
Section # _____________________ Division: ______ SS _____PS/C
Competitor # __________________ Judge’s Signature ______________________
*For all Judge verification steps, full points are only awarded if all components are accurate
Skill II: Set Up Restriction Digestion Reaction (Time: 15 minutes)
Possible
Awarded
1. Entered testing area wearing closed-toed shoes and donned proper PPE:
glasses/safety glasses/goggles, and gloves (lab coat is optional).
2 0
2. Labeled 6 microcentrifuge tubes to match the labels on the DNA samples provided.
*Judge verified competitor labeled all 6 tubes correctly.
2 0
3. Pipetted 10 µl of each DNA sample into the corresponding labeled microcentrifuge
tube.
*Judge verified competitor (i) used a 20 µl micropipet, (ii) set it to 10 µl, (iii) used an
appropriate and clean pipet tip for the sample, and (iv) transferred liquid without air
bubbles or losing sample in the transfer process.
4 0
4. Pipetted 10 µl of enzyme mix (ENZ) into each PCR tube then mixed by pipetting
up and down 2-3 times.
*Judge verified competitor (i) used a 20 µl micropipet set to 10 µl, (ii) used a clean and
appropriate pipet tip between each sample, and (iii) transferred liquid without air bubbles
or losing sample in the transfer process.
4 0
5. Capped each tube tightly and mixed by flicking each tube with fingers.
2 0
6. Pulse-spinned tubes in a microcentrifuge -or- tapped tubes on table to collect all
liquid at the bottom of the tube.
*Judge verified competitor balanced the tubes in the microcentrifuge or tapped them on
the table to collect liquid.
4 0
7. Verbalized one of the two options for incubation: (i) incubating reactions at room
temperature overnight -or – (ii) incubating reactions at 37⁰C for 45 min.
4 0
Cleaning up
8. Cleaned work area:
a. Disposed of pipet tips and microcentrifuge tubes in waste receptacle.
2 0
b. Cleaned work area with surface disinfectant.
2 0
c. Removed PPE.
2 0
d. Washed hands or used alcohol-based hand-rub for hand hygiene.
*Judge verified steps 8a-d were performed in the order written here.
2 0
TOTAL POINTS - SKILL II
70% Mastery for Skill II = 21.0
30
HOSA Biotechnology Event Guidelines (August 2023) Page 8 of 17
Biotechnology
Section # _____________________ Division: ______ SS _____PS/C
Competitor # __________________ Judge’s Signature ______________________
*For all Judge verification steps, full points are only awarded if all components are accurate
Skill III: DNA Gel Electrophoresis (Time: 20 minutes)
Possible
Awarded
1. Entered testing area wearing closed-toed shoes and donned proper PPE:
glasses/safety glasses/goggles, and gloves (lab coat is optional).
2 0
Preparing samples
2. Collected the liquid to the bottom of the 6 samples and DNA size standard by either
(i) placing tubes into microcentrifuge or mini centrifuge and pulse-spinning for 5–10
seconds, or (ii) by tapping the tubes gently on the table.
*Judge verified competitor balanced the tubes in the microcentrifuge or tapped them on
the table to collect liquid.
4 0
3. Pipetted 5 µl of sample loading buffer (SLB) into each tube. Pipetted up and down or
flicking the tubes to mix.
*Judge verified competitor (i) used 20 µl micropipet w proper pipet tip, (ii) set micropipet to
deliver 5 µl to all 7 tubes, (iii) used a fresh pipet tip for each sample, and (iv) transferred
liquid without air bubbles or losing sample in the transfer process.
4 0
Loading the gel
4. Placed the precast agarose gel into the electrophoresis chamber.
*Judge verified competitor placed the wells of the agarose gel near the black (-) electrode
(cathode).
4 0
5. Filled the electrophoresis chamber with 1x TAE buffer; added enough buffer to
cover the gel and fill the wells.
2 0
6. Loaded 10 µl standard into a well in the gel.
*Judge verified competitor (i) selected 20 µl micropipet and set it to deliver 10 µl, (ii) used a
fresh pipet tip of the correct type, and (iii) loaded standard into the gel with no gel
breakage or sample overflow into nearby wells.
4 0
7. Loaded 20 µl of each sample into separate wells of the gel.
*Judge verified competitor (i) selected 20 µl or 200 µl micropipet and set it to deliver 20 µl,
(ii) used a fresh pipet tip of the correct type for each sample, and (iii) loaded samples into
the gel with no gel breakage, piercing the bottom of wells, or sample overflow into nearby
wells.
4 0
8. Recorded the order of sample loading on the sheet provided.
2 0
9. Placed the lid on the electrophoresis chamber and connected the electrical leads to
the power supply.
*Judge verified competitor connected red to red and black to black.
1 0
10. Turned on the power and ran the gel at 100 V.
*This step may be verbalized.
2 0
11. Verbalized that would run for 30 minutes.
2 0
12. Verbalized run has completed, then turned off power and removed lid from
chamber.
1 0
HOSA Biotechnology Event Guidelines (August 2023) Page 9 of 17
Skill III: DNA Gel Electrophoresis (cont.)
Possible
Awarded
Cleaning up
13. Cleaned work area:
a. Disposed of pipet tips, microcentrifuge tubes, and gel in waste receptacle.
2 0
b. Cleaned work area with surface disinfectant.
2 0
c. Removed PPE.
2 0
d. Washed hands or used alcohol-based hand-rub for hand hygiene.
*Judge verified steps 13a-d were performed in the order written here.
2 0
TOTAL POINTS - SKILL III
70% Mastery for Skill III = 28
40
HOSA Biotechnology Event Guidelines (August 2023) Page 10 of 17
Biotechnology
Section # _____________________ Division: ______ SS _____PS/C
Competitor # __________________ Judge’s Signature ______________________
Skill IV: DNA Gel Interpretation (Time: 15 minutes)
Possible
Awarded
1. Using a ruler, measured the distance (in mm) that each of the DNA fragments or
bands traveled from the well. Recorded results for each sample and standard in the
table provided.
4 0
2. Using the semilog graph paper provided, plotted the distance versus size for the
bands in the standard.
6 0
3. Drew a line of best fit through the points.
4 0
4. Used the graph to estimate the fragment size for each band in the samples.
Recorded estimates in the table provided.
6 0
TOTAL POINTS - SKILL IV
70% Mastery for Skill IV = 14
20
HOSA Biotechnology Event Guidelines (August 2023) Page 11 of 17
Biotechnology
Section # _____________________ Division: ______ SS _____PS/C
Competitor # __________________ Judge’s Signature ______________________
*For all Judge verification steps, full points are only awarded if all components are accurate
Skill V: Bradford Protein Quantitation Assay (Time: 20 minutes)
Possible
Awarded
1. Entered testing area wearing closed-toed shoes and donned proper PPE:
glasses/safety glasses/goggles, and gloves (lab coat is optional).
2 0
Preparing samples
2. Labeled two empty microcentrifuge tubes for the sample dilutions (for example, 1/50
A and 1/50 B.
1 0
3. Pipetted 2 µl of sample A into the microcentrifuge tube labeled 1/50 A.
*Judge verified competitor (i) selected 20 µl micropipet, (ii) set micropipet to correct
volume, (iii) used a clean pipet tip, (iv) used a proper pipet tip, and (v) transferred liquid
without air bubbles or losing sample in the transfer process.
2 0
4. Pipetted 98 µl of 1x PBS into the microcentrifuge tube labeled 1/50 sample A.
Judge verified competitor (i) selected (20-200 µl micropipet, (ii) set micropipet to correct
volume, (iii) used a clean pipet tip, (iv) used a proper pipet tip, and (v) transferred liquid
without air bubbles or losing sample in the transfer process.
2 0
5. Mixed well by pipetting, flicking, or vortexing.
2 0
6. Pipetted 2 µl of sample B into the microcentrifuge tube labeled 1/50 B.
*Judge verified competitor (i) selected 20 µl micropipet, (ii) set micropipet to correct
volume, (iii) used a clean pipet tip, (iv) used a proper pipet tip, and (v) transferred liquid
without air bubbles or losing sample in the transfer process.
2 0
7. Pipetted 98 µl of 1x PBS into the microcentrifuge tube labeled 1/50 sample B.
Judge verified competitor (i) selected 20-200 µl micropipet, (ii) set micropipet to correct
volume, (iii) used a clean pipet tip, (iv) used a proper pipet tip, and (v) transferred liquid
without air bubbles or losing sample in the transfer process.
2 0
8. Mixed well by pipetting, flicking, or vortexing.
2 0
9. Labeled two cuvettes: Sample A and Sample B (or just A and B).
1 0
10. Pipetted 20 µl of the 1/50 diluted samples into the corresponding cuvettes.
*Judge verified competitor (i) selected 20 µl micropipet, (ii) set micropipet to correct
volume, (iii) used a clean pipet tip for each liquid, (iv) used a proper pipet tip, and (v)
transferred liquid without air bubbles or losing sample in the transfer process.
4 0
Preparing standards
11. Labeled eight cuvettes for the protein standards: Blank, 0.125, 0.250, 0.500,
0.750, 1.000, 1.500, 2.000
1 0
12. Pipetted 20 µl of 1x PBS into the cuvette labeled Blank.
*Judge verified competitor (i) selected correct micropipet (20 µl), (ii) set micropipet to
correct volume, (iii) used a clean pipet tip, (iv) used a proper pipet tip, and (v) transferred
liquid without air bubbles or losing sample in the transfer process.
4 0
HOSA Biotechnology Event Guidelines (August 2023) Page 12 of 17
Skill V: Bradford Protein Quantitation Assay (cont.)
Possible
Awarded
13. Pipetted 20 µl of each protein standard into a corresponding cuvette.
*Judge verified competitor (i) selected correct micropipet (20 µl), (ii) set micropipet to
correct volume, (iii) used a clean pipet tip for each standard, (iv) used a proper pipet tip,
and (v) transferred liquid without air bubbles or losing sample in the transfer process.
4 0
Adding Bradford reagent
14. Added 1 ml of the 1x Bradford reagent to all ten cuvettes. Mixed well by pipetting
up and down.
*Judge verified competitor (i) used the correct micropipet (100-1,000 µl), (ii) set
micropipet to correct volume, (iii) used a fresh pipet tip for each sample, (iv) used a
proper pipet tip, (v) transferred liquid without air bubbles or losing sample in the
transfer process, and (vi) pipetted up and down gently to mix (no liquid sucked into the
barrel of the micropipet, for example).
4 0
15. Verbalized that cuvettes would incubate at room temperature for 5 minutes.
1 0
16. After visually comparing the cuvettes containing samples to the cuvettes containing
the protein standard, verbalized an estimated protein concentration of the samples.
4 0
Cleaning up
17. Cleaned work area:
a. Disposed of pipet tips, microcentrifuge tubes, and cuvettes in waste
receptacle.
2 0
b. Cleaned work area with surface disinfectant.
2 0
c. Removed PPE.
2 0
d. Washed hands or used alcohol-based hand-rub for hand hygiene.
*Judge verified steps 17a-d were performed in the order written here.
2 0
TOTAL POINTS - SKILL V
70% Mastery for Skill V = 32.2
46
HOSA Biotechnology Event Guidelines (August 2023) Page 13 of 17
Biotechnology
Section # _____________________ Division: ______ SS _____PS/C
Competitor # __________________ Judge’s Signature ______________________
*For all Judge verification steps, full points are only awarded if all components are accurate
Skill VI: Bacterial Transformation (Time: 20 minutes)
Possible
Awarded
1. Entered testing area wearing closed-toed shoes and donned proper PPE:
glasses/safety glasses/goggles, and gloves (lab coat is optional).
2 0
Preparing for heat shock
2. Labeled one microcentrifuge tube +pGLO and another -pGLO.
1 0
3. Pipetted 250 µl of transformation solution (0.05 M CaCl
2
) into each tube, placed
tubes on ice.
*Judge verified competitor (i) selected 100-1,000 µl micropipet, (ii) set it to deliver 250 µl,
(iii) used a proper pipet tip, and (iv) transferred liquid without air bubbles or losing sample
in the transfer process.
4 0
4. Used a sterile plastic inoculation loop to scrape 2–4 E. coli colonies from the
surface of the starter plate.
2 0
5. Transferred the loop into the +pGLO tube and swirled it in the transformation
solution to disperse bacteria. Closed the tube and placed it back on ice.
2 0
6. Disposed of loop into biohazard waste receptacle or onto a paper towel for disposal
later.
2 0
7. Used a second sterile plastic inoculation loop to scrape 2–4 E. coli colonies from the
surface of the starter plate.
2 0
8. Transferred the loop into the -pGLO tube and swirled it in the transformation
solution to disperse bacteria.
2 0
9. Disposed of loop into biohazard waste receptacle or onto a paper towel for disposal
later.
2 0
10. Pipetted 10 µl of pGLO plasmid into the +pGLO tube and mixed by pipetting
gently up and down.
*Judge verified competitor (i) used 20 µl micropipet and set it to deliver 10 µl using a
correct tip and (ii) did NOT add plasmid to the -pGLO tube.
4 0
11. Disposed of pipet tip into biohazard waste receptacle or onto a paper towel for
disposal later.
2 0
12. Placed both tubes back on ice, making sure the tubes were in full contact with the
ice.
1 0
13. Labeled agar plates with “+pGLO” or “-pGLO” as follows:
LB/amp +pGLO
LB/amp/ara +pGLO
LB/amp -pGLO
LB -pGLO
*Judge verified competitor labeled the bottoms of the plates and not the lids.
4 0
14. Verbalized a 10 min incubation on ice had completed.
2 0
HOSA Biotechnology Event Guidelines (August 2023) Page 14 of 17
Skill VI: Bacterial Transformation (cont.)
Possible
Awarded
Performing heat shock
15. Transferred the +pGLO and -pGLO tubes from the ice into a 42
0
C water bath for
exactly 50 seconds making sure the tubes were in full contact with the water.
Immediately placed tubes back on ice.
4 0
16. Verbalized that the tubes remained on ice for 2 minutes.
2 0
17. Removed the tubes from ice and pipetted 250 µl of LB broth into each tube.
*Judge verified competitor (i) used 100-1,000 µl micropipet to deliver 250 µl and (ii)
changed the pipet tip between each sample, (iii) used a proper pipet tip, and (iv)
transferred liquid without air bubbles or losing sample in the transfer process.
4 0
18. Disposed of pipet tips into biohazard waste receptacle or onto a paper towel for
disposal later.
2 0
19. Verbalized the samples incubated at room temperature for 10 min.
1 0
Plating the bacteria
20. Mixed the tubes by inverting or flicking.
1 0
21. Pipetted 100 µl of the transformation mixtures onto appropriately labeled agar
plates (for example, a “+pGLO” mixture onto a “+pGLO plate”, etc.).
*Judge verified competitor (i) used 20-200 µl micropipet to deliver 100 µl to each plate, (ii)
applied correct sample to correct plate, (iii) changed the pipet tip between each sample,
(iv) used a proper pipet tip, (v) transferred liquid without air bubbles or losing sample in
the transfer process, and (vi) applied correct sample to correct plate.
4 0
22. Disposed of pipet tips into biohazard waste receptacle or onto a paper towel for
disposal later.
2 0
23. Used a sterile plastic inoculation loop to spread the bacteria over the entire
surface of the plate in all directions. Disposed of loop into biohazard waste
receptacle or onto a paper towel for disposal later.
4 0
24. Repeated step 23 for each plate.
*Judge verified competitor used a new loop for each plate and the samples matched plates.
10 0
25. Stacked plates together with lids facing downward, agar side facing up.
*Judge verified competitor placed plates with agar side up.
4 0
26. Verbalized plates would incubate at 37
0
C for 16–24 hours.
2 0
Cleaning up
27. Cleaned work area:
a. Disposed of pipet tips, microcentrifuge tubes, and loops into biohazard waste
receptacle, cleaned area of any spilled liquid.
2 0
b. Cleaned work area with surface disinfectant.
2 0
c. Removed PPE.
2 0
d. Washed hands or used alcohol-based hand-rub for hand hygiene.
*Judge verified steps 27a-d were performed in the order written here.
2 0
TOTAL POINTS - SKILL VI
70% Mastery for Skill VI = 56
80
HOSA Biotechnology Event Guidelines (August 2023) Page 15 of 17
Biotechnology
Section # _____________________ Division: ______ SS _____PS/C
Competitor # __________________ Judge’s Signature ______________________
*For all Judge verification steps, full points are only awarded if all components are accurate
Skill VII: Calculation of Transformation Efficiency (Time: 10 minutes)
Possible
Awarded
1. Entered testing area wearing closed-toed shoes and donned proper PPE:
glasses/safety glasses/goggles, and gloves (lab coat is optional).
2 0
2. Counted the number of transformed colonies on the plate and recorded that number
on the printed scenario or paper provided. If there are >~50 colonies, an estimation
made by counting colonies in a quadrant on the plate is acceptable.
4 0
3. Calculated how many micrograms of DNA were spread onto the plate.
4 0
4. Expressed answer to #3 using correct units (µg).
2 0
5. Calculated the transformation efficiency.
4 0
6. Expressed answer to #5 using correct units (CFU/µg or colonies/µg).
2 0
7. Removed PPE before leaving the area.
2 0
TOTAL POINTS - SKILL VII
70% Mastery for Skill VII = 14
20
HOSA Biotechnology Event Guidelines (August 2023) Page 16 of 17
Biotechnology
Section # _____________________ Division: ______ SS _____PS/C
Competitor # __________________ Judge’s Signature ______________________
*For all Judge verification steps, full points are only awarded if all components are accurate
Skill VIII: Qualitative ELISA (Time: 20 minutes)
Possible
Awarded
1. Entered testing area wearing closed-toed shoes and donned proper PPE:
glasses/safety glasses/goggles, and gloves (lab coat is optional).
2 0
2. Labeled a 12-well microplate strip:
• the first three wells with + for the positive controls
• the next three wells with a – for the negative control
• the next three wells with an S to indicate the sample.
2 0
Antigen incubation
3. Transferred 50 µl of purified antigen (AG) into each well.
*Judge verified competitor added AG to all 9 wells and (i) used a 20-200 µl micropipet, (ii)
set it to 50 µl, (ii) used a proper pipet tip, and (iii) transferred liquid without air bubbles or
losing sample in the transfer process.
4 0
4. Incubated the samples at room temperature for 2 min.
2 0
5. Followed wash protocol:
a. Tipped each microplate strip upside-down onto a short stack of paper
towels and gently tapped strip a few times to drain the wells while making
sure to avoid splashing sample back into wells.
2 0
b. Discarded the wet paper towels.
2 0
c. Used a transfer pipet (same transfer pipet can be used) to fill each well with
wash buffer, taking care not to touch the well or spill the buffer into
neighboring wells.
2 0
6. Repeated step #5.
2 0
Sample incubation (primary antibody)
7. Transferred 50 µl of the positive control (+) into the three + wells.
*Judge verified competitor added + to only the first 3 wells labeled + and (i) used a 20-200
µl micropipet set to 50 µl, (ii) used a proper pipet tip, and (iii) transferred liquid without air
bubbles or losing sample in the transfer process.
4 0
8. Transferred 50 µl of the negative control (-) into the three – wells.
*Judge verified competitor added - to only the 3 wells labeled - and (i) used a 20-200 µl
micropipet set to 50 µl, (ii) used a proper pipet tip, and (iii) transferred liquid without air
bubbles or losing sample in the transfer process.
4 0
9. Transferred 50 µl of the sample (S) into the corresponding three wells.
*Judge verified competitor added S to only the 3 wells labeled S and (i) used a 20-200 µl
micropipet set to 50 µl, (ii) used a proper pipet tip, and (iii) transferred liquid without air
bubbles or losing sample in the transfer process.
4 0
10. Incubated the samples at room temperature for 2 minutes.
1 0
11. Performed wash protocol (step 5) two times.
2 0
HOSA Biotechnology Event Guidelines (August 2023) Page 17 of 17
Skill VIII: Qualitative ELISA (cont.) (Time: 20 minutes)
Possible
Awarded
Enzyme-linked antibody (secondary antibody) incubation
12. Transferred 50 µl of enzyme-linked antibody (ELA) into each well.
*Judge verified competitor added ELA to all 9 wells and (i) used a 20-200 µl micropipet set
to 50 µl, (ii) used a proper pipet tip, and (iii) transferred liquid without air bubbles or losing
sample in the transfer process.
4 0
13. Incubated the samples at room temperature for 2 minutes.
1 0
14. Performed wash protocol (step 5) three times.
2 0
Substrate incubation and color development
15. Transferred 50 µl of enzyme substrate (SUB) into each well.
*Judge verified competitor added SUB to all 9 wells and (i) used a 20-200 µl micropipet set
to 50 µl, (ii) used a proper pipet tip, and (iii) transferred liquid without air bubbles or losing
sample in the transfer process.
4 0
Cleaning up
16. Cleaned work area:
a. Disposed of pipet tips, microcentrifuge tubes, transfer pipets, and paper towels
into waste receptacle, cleaned area of any spilled liquid, returned micropipets
to rack (if available).
2 0
b. Cleaned work area with surface disinfectant.
2 0
c. Removed PPE.
2 0
d. Washed hands or used alcohol-based hand-rub for hand hygiene.
*Judge verified steps 16a-d were performed in the order written here.
2 0
17. Observed and reported results.
*Judge verified (+) and S wells were blue, (-) was colorless; competitor confirmed sample
was positive.
4 0
TOTAL POINTS – SKILL VI
70% Mastery for Skill VI = 39.2
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