Protein-purification lab
When you actually run the column today, which of your numbered fractions will hold the target , and can you prove it with the green glow under UV?
Run a - procedure and collect fractions to isolate the target protein.
- • You'll be able to run a column and collect fractions.
- • You'll be able to identify target fractions using the GFP signal.
- Before you load the column, predict: will the target come out in the early wash fractions or the later fractions? Explain in one line.
- What single piece of evidence today will tell you a fraction contains your target ?
- 1Read the lab protocol in the PLTW course shell and set up your column and tubes.
- 2Load the mixture and begin collecting numbered fractions.
- 3Add buffer to elute the bound and watch for the GFP signal under UV.
- 4Record which fractions glow and label them as your target collection.
- 5Note one source of error and how you controlled for it.
- 6Submit your fraction-collection data sheet.
🛠 Get unstuck · pick your level
Lab day: Tier 1 is the whole class at the bench. No extension today.
🔑 Today's words · 5
Tap a word in the lesson for a plain meaning and one example. Recycled into next week's Do-Now.
Do the work · 80-minute blockfirst 5 min = hook▸
💡 Big idea: Column physically isolates the target because only it binds the resin tightly, so contaminants wash out first and the target elutes last into the glowing fractions.
- 0-10Read protocol; set up column and label collection tubes
- 10-25Load mixture; begin collecting fractions as wash proceeds
- 25-45Add buffer; collect elution fractions in order
- 45-58Check fractions under UV; record which fractions glow
- 58-70Label target fractions; note source of error and control
- 70-80Clean up column and bench; submit fraction-collection data sheet
- • Today is the - lab.
- • You will run a column, collect numbered fractions, and use UV light to find your .
- • Work precisely: fraction order matters, and mislabeling a tube loses data you cannot recover.
- • Fraction data collected today feeds directly into Thursday's gel interpretation.
- • Loading the lysate applies all proteins to the resin; only the target binds with high affinity.
- • Wash steps remove loosely bound contaminants before begins.
- • GFP fluorescence under UV is the qualitative signal that confirms the target is in a fraction.
GFP, chromatography, SDS-PAGE / gel interpretation, purity and QC. · - lab
Day 3 of this lesson. Open this exact section in myPLTW (find it in Clever, Microsoft sign-in), then do the work below.
Do this: Open Activity 4.1.3 GFP in myPLTW and run the protein-purification lab protocol to collect and identify your target fractions.
Mark the - lab entry complete and attach your fraction-collection data sheet.
diagram should be done (Tuesday); fraction-collection data sheet due today.
Fraction-collection data sheet with tube numbers, buffer, GFP signal, and error-control note submitted.
All PLTW activities are completed inside the PLTW course environment: this page only gives direction. Submit producibles on Schoology.
Check things off as you work, then submit. This tells Mr. Mendoza how you're doing so he can help the class. It does not replace turning in your producible on Schoology.
Use the code Mr. Mendoza gave you, not your name. Saved on this device.
GFP, chromatography, SDS-PAGE / gel interpretation, purity and QC. · Protein-purification lab
Open Activity 4.1.3 GFP in myPLTW and run the protein-purification lab protocol to collect and identify your target fractions.
diagram should be done (Tuesday); fraction-collection data sheet due today.
This is how Mr. Mendoza sees the class keeping pace with PLTW. Be honest, it only helps if it is accurate.
🎯 Run a - procedure and collect fractions to isolate the target protein.
- Read the lab protocol in the PLTW course shell and set up your column and tubes.
- Load the mixture and begin collecting numbered fractions.
- Add buffer to elute the bound and watch for the GFP signal under UV.
- Record which fractions glow and label them as your target collection.
- Note one source of error and how you controlled for it.
- Submit your fraction-collection data sheet.
Data table: Fraction-collection data sheet recording tube number, buffer applied, GFP signal (yes/no), and target-fraction labels plus one error-control note.
Turn it in on Schoology using the checklist just below. Upload by 11:29 PM for full credit.
| Task | Who |
|---|---|
| Read the lab protocol in the PLTW course shell and set up your column and tubes. | _______ |
| Load the mixture and begin collecting numbered fractions. | _______ |
| Add buffer to elute the bound and watch for the GFP signal under UV. | _______ |
| Record which fractions glow and label them as your target collection. | _______ |
| Note one source of error and how you controlled for it. | _______ |
| Submit your fraction-collection data sheet. | _______ |
Working solo? Put your own name in "Who" for every row.
- You'll be able to run a column and collect fractions.
- You'll be able to identify target fractions using the GFP signal.
- 1Do thisRun a protein-purification procedure and collect fractions to isolate the target protein.
- 2Use this resource
- 3Submit thisData table: Fraction-collection data sheet recording tube number, buffer applied, GFP signal (yes/no), and target-fraction labels plus one error-control note.
- 4Submit it here
- 1Open Clever.
- 2Microsoft (district) sign-in.
- 3Schoology and myPLTW are both in Clever.
Look for this assignment in Schoology: Genetics of Disease (Medical Interventions) › GFP, chromatography, SDS-PAGE / gel interpretation, purity and QC. › Data tableOpen Schoology
Learn it · deck, reading, and vocabulary▸
Tier 1 is the time-boxed teacher set for the block; Tier 2 adds scaffolded vocabulary, examples, and a reading routine; Tier 3 extends into careers and current biomedical applications.
Generated from this lesson's canonical data with a red-team citation check.
Students often think Students expect the target to drip out first and early, since it was loaded first, so they watch the wrong tubes.. The trap: The target binds the resin and stays put through the loading and wash steps while contaminants leave first. It only comes out during , in the later fractions, so the glow shows up after the wash, not at the start.
I loaded the protein mixture, washed, then eluted while watching for the green GFP signal under UV. Here is my record.
Reading: tubes 1 and 2 were the flow-through and wash, with no glow, meaning contaminants washed off. Tubes 4 and 5 glowed green, so those are my target collection.
Error-control note: one source of error is room light washing out the faint UV glow, so I controlled for it by reading each tube in the same darkened spot with the same UV lamp distance, so my yes/no calls are consistent.
| Tube | Buffer applied | GFP signal | Label |
|---|---|---|---|
| 1 | Load flow-through | No | Discard |
| 2 | Wash | No | Discard |
| 4 | Elution | Yes | Target |
| 5 | Elution | Yes | Target |
Also due today: Submit your fraction-collection data sheet to the course shell before leaving lab.
- CER:
- Claim, Evidence, Reasoning: make a claim, back it with evidence, explain your reasoning.
- SOP:
- Standard Operating Procedure, the exact steps to follow (especially in a lab).
- Tracker:
- Your PLTW progress log where you record completed evidence.
- myPLTW:
- The PLTW course site where you do the online activities. Find it in Clever with your Microsoft sign-in, right next to Schoology.
Tap the speaker to hear a term. Add two of these to your notebook glossary with a definition and an example in your own words.
Pick just 2 or 3 words from today and make them yours: write what each one means in your own words, then give one example from what you actually did in Protein-purification lab. Try your own words first; the glossary is there if you get stuck. This is voluntary and counts as extra credit, so keep it short.
Saved on this device. Show Mr. Mendoza or add these to your notebook glossary to claim the extra credit.
Classroom documents for this lesson are posted in Schoology. Open Schoology and find each one by the name shown on its card.
Open this when the class reaches this activity and use it to complete the required lesson artifact.
Placement rationale
Matched and quality control by path:Medical-Interventions/Unit-4_When-Organs-Fail/4.1_Manufacturing-Human-Proteins; keywords:protein purification, gfp, . Score 150. Visibility: student-schoology (student-facing resource; link through Schoology rather than local path).
Open this when the class reaches this activity and use it to complete the required lesson artifact.
Placement rationale
Matched and quality control by path:Medical-Interventions/Unit-4_When-Organs-Fail/4.1_Manufacturing-Human-Proteins; keywords:protein purification, gfp, . Score 150. Visibility: student-schoology (student-facing resource; link through Schoology rather than local path).
Use this if you were absent, got stuck, or need another pass before you submit the lesson artifact.
Placement rationale
Matched and quality control by path:Medical-Interventions/Unit-4_When-Organs-Fail/4.1_Manufacturing-Human-Proteins; keywords:gfp, . Score 142. Visibility: student-schoology (student-facing resource; link through Schoology rather than local path).
How to get there: open Clever and sign in with your Microsoft (district) account. You will find both Schoology and myPLTW right there in Clever. Turn in your work on Schoology; do the online activities in myPLTW.
Check yourself · commit, then reveal▸
Your fractions 1-5 are collected during the wash and stay dark under UV; fractions 8 and 9 glow green. Which fractions are your target, and why did the target not appear in fractions 1-5?
Write an answer and pick a confidence to unlock the key.
Fast retrieval with instant answers, not the commit-then-reveal check above. Try each from memory first: write what you remember about the earlier units, then check yourself here.
Go further and get help▸
Finish the checklist before you handle any material.
- • Wear nitrile gloves and safety goggles throughout the procedure.
- • UV light is harmful to eyes and skin; never look directly into the UV lamp and minimize exposure to skin.
- • Treat all protein samples as potential allergens; avoid contact with eyes and mouth.
- • Dispose of all biological waste in designated containers per lab protocol.
- • Wipe the bench with 70% ethanol before and after the procedure.
- • Report any spill or broken glassware to the teacher immediately.
- 1Frame the guiding question and name your independent and dependent variables.
- 2Plan a method that would actually answer it, then get the plan checked before you start.
- 3Collect data carefully and record exactly what you observe before you interpret it.
- 4Build a tentative argument on a whiteboard: claim, evidence, reasoning.
- 5Argumentation session: present your board, question another group, and revise your claim.
- 6Write the final CER with your strongest evidence and one named limitation of the method.
What today's skills lead to. These are real health-science careers this course builds toward. Tap one to see, on the US Department of Labor's O*NET site, what the job actually involves, what it pays, and how fast it is growing.
Complete the virtual - run linked in the course shell, recording which fractions show the GFP signal under UV, then submit your fraction-collection data sheet.
Learn.Genetics: Gel ElectrophoresisThen submit your Data table on Schoology.
Class still runs. Complete the online activity above (it's self-guided). Need the concept taught without a teacher? Use this authoritative explainer:
Genetic Science Learning Center: Genetics basics and proteinsYou've passed Unit 2, so the optional extra-credit track is open. Complete reserved-unit work from home (virtual labs included) for extra credit, submitted on Schoology.
Open the extra-credit track- CompleteEvery required part of the artifact is present, nothing left blank.
- AccurateThe science and the data are correct and match the evidence.
- Scientific reasoningYou explain your claim with evidence and reasoning (CER), not just an answer.
- Professional communicationClear, organized, labeled, and written the way a clinician or scientist would.
- SubmittedTurned in the right way (Schoology for routine work) and confirmed.
- Error analysis and method · counts doubleName a specific limit of the method and how it moved your result, and compare what you predicted to what happened. "Human error" does not count; say what about the procedure or instrument caused it.