Cloning and purification workflow
Safety gate · before any work
- Wear nitrile gloves and safety goggles throughout the procedure.
- Treat all bacterial cultures as BSL-1 organisms: avoid mouth contact and wash hands before leaving lab.
- Dispose of all biological waste (plates, tubes, tips) in designated biohazard bags.
Do now
Carry out a cloning and protein-purification workflow and record results at each step.
- Hand in
- Cloning and purification workflow data table recording transformation results, selection counts, fraction data, yield, and a quality observation.
- Where
- Turn this in at the drop folder with your district Microsoft sign-in, or hand it to Mr. Mendoza in class. Both count as submitted. Doing the activity in myPLTW does not.
You get two school days for every day you were absent, so this deadline moves with you.
You built on paper yesterday, but how do you force millions of bacteria to actually swallow that , and how do you find the few that did?
Carry out a cloning and - and record results at each step.
- • You'll be able to carry out , selection, and steps.
- • You'll be able to record yield and a quality note from your .
- Bacteria do not normally let loose DNA cross their membranes. Name one physical way you might force a membrane to briefly open.
- If you mix DNA with a billion bacteria and only some take it up, how could you kill off the ones that failed, so only the successful cells are left?
- 1Read the protocol in the PLTW course shell and gather your materials.
- 2Model by introducing the recombinant into host cells.
- 3Select transformed cells using the provided marker and record how many grew.
- 4Run the simulated step and note where the target appears.
- 5Record yield and one quality observation in your .
- 6Submit your cloning and results.
What did this day actually feel like?
Cloning and purification workflow
LAB The whole workflow end to end: cut, insert, transform, select, express, purify. Every step has a way to fail and a way to check.
Turned in: lab report → Lab Reports folder
Fiction. There is no such student. The lessons, labs and dates are the real planned course; the student, the classmates and the conversations are invented.
The same day, drawn.
The whole workflow end to end: cut, insert, transform, select, express, purify. Every step has a way to fail and a way to check.
Fiction. There is no such student. The lessons, labs and dates are the real planned course; the student, the classmates and the conversations are invented.
🛠 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: A recombinant can be mass-produced because puts the into cells, selection keeps only the cells that took it, and separates the protein from debris, so each step sets up the next.
- 0-10Review protocol; gather materials; confirm lab setup
- 10-30 step: introduce into host cells per protocol
- 30-45Selection: plate or score cells; count or estimate transformed colonies
- 45-60 step: run first ; identify fraction with target
- 60-72Record yield and quality observation in
- 72-80Clean up; submit to the class site
- • This is a hands-on bacterial lab.
- • You will introduce into host bacteria, select for transformed cells, and run a first step.
- • Work carefully: at any step ruins results.
- • Lab SOPs and data recording are both scored domains on the 072130 WebXam.
- • Heat shock or electroporation opens pores in bacterial membranes so DNA can enter.
- • selection kills non-transformed cells; only cells carrying the resistance gene survive.
- • The first step separates soluble from cell debris before .
Plasmids, restriction enzymes, ligase, transformation, protein expression. · Cloning and
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 4.1.2 Factories in myPLTW and follow the and selection protocol to model the cloning and .
Mark the cloning- entry complete and attach your workflow .
Cloning-tools diagram should be done (Tuesday); due today.
Cloning and with results, selection counts, and fraction data submitted.
The official PLTW activity stays inside myPLTW. If myPLTW will not open, use F1 and E1-E3 on this page to complete today's local evidence decision, then make up the official activity when access returns. Turn this in at the drop folder with your district Microsoft sign-in, or hand it to Mr. Mendoza in class. Both count as submitted. Doing the activity in myPLTW does not.
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 through the submission route shown below.
Use the code Mr. Mendoza gave you, not your name. Saved on this device.
Plasmids, restriction enzymes, ligase, transformation, protein expression. · Cloning and purification workflow
Open Activity 4.1.2 Factories in myPLTW and follow the and selection protocol to model the cloning and .
Cloning-tools diagram should be done (Tuesday); due today.
This is how Mr. Mendoza sees the class keeping pace with PLTW. Be honest, it only helps if it is accurate.
🎯 Carry out a cloning and - and record results at each step.
- Read the protocol in the PLTW course shell and gather your materials.
- Model by introducing the recombinant into host cells.
- Select transformed cells using the provided marker and record how many grew.
- Run the simulated step and note where the target appears.
- Record yield and one quality observation in your .
- Submit your cloning and results.
Lab report: Cloning and recording results, selection counts, fraction data, yield, and a quality observation.
Turn this in at the drop folder with your district Microsoft sign-in, or hand it to Mr. Mendoza in class. Both count as submitted. Doing the activity in myPLTW does not. Use the checklist just below and upload by 11:29 PM for full credit. Absent with an excused absence? You get two school days for every day you were absent, so this deadline moves with you.
| Task | Who |
|---|---|
| Read the protocol in the PLTW course shell and gather your materials. | _______ |
| Model by introducing the recombinant into host cells. | _______ |
| Select transformed cells using the provided marker and record how many grew. | _______ |
| Run the simulated step and note where the target appears. | _______ |
| Record yield and one quality observation in your . | _______ |
| Submit your cloning and results. | _______ |
Working solo? Put your own name in "Who" for every row.
- You'll be able to carry out , selection, and steps.
- You'll be able to record yield and a quality note from your .
- 1Do thisCarry out a cloning and protein-purification workflow and record results at each step.
- 2Use this resource
- 3Submit thisLab report: Cloning and purification workflow data table recording transformation results, selection counts, fraction data, yield, and a quality observation.
- 4Submit it here
- 1Open the drop folder.
- 2Sign in with your district Microsoft account, not a personal one.
- 3Upload the file, named Lastname_Firstname__Assignment Title.
- 4Your own upload panel says Uploaded with a green check: that is your receipt.
Turn this in at the drop folder with your district Microsoft sign-in, or hand it to Mr. Mendoza in class. Both count as submitted. Doing the activity in myPLTW does not. Genetics of Disease (Medical Interventions) › Plasmids, restriction enzymes, ligase, transformation, protein expression. › Lab reportOpen the drop folder
Learn it · deck, reading, and vocabulary▸
The deck carries the prior idea forward, lets you inspect an analogy, maps the rule to biology, and ends with the same evidence decision and exit ticket used on this page.
Generated from this lesson's canonical data with a red-team citation check.
Life-saving biotechnology raises hard questions about access because the same product must pass separate gates of price, patents, and regulation, so who controls those gates decides who the science actually helps.
A recombinant can be mass-produced because puts the into cells, selection keeps only the cells that took it, and separates the protein from debris, so each step sets up the next.
A library keeps a master plan protected while working copies guide production at different stations.
- Why protect the master copy?
- What information moves?
- Where can an error change the final product?
Stored information can be copied, read, and converted into a functional product.
Genes are regulated biological sequences, not conscious instructions, and one gene rarely determines a whole trait alone.
- • Master plan maps to DNA.
- • Working copy maps to RNA.
- • Production output maps to or a regulated cell function.
Driving question: You built on paper yesterday, but how do you force millions of bacteria to actually swallow that , and how do you find the few that did?
What you already know: Life-saving biotechnology raises hard questions about access because the same product must pass separate gates of price, patents, and regulation, so who controls those gates decides who the science actually helps.
New idea: A recombinant can be mass-produced because puts the into cells, selection keeps only the cells that took it, and separates the protein from debris, so each step sets up the next.
Visual or model: F1. F1. A lesson illustration or teaching diagram for Cloning and purification workflow. Use it with E1-E3; it is a model or context image, not experimental or patient data. What to notice: Trace the labeled testing, treatment, or biological process and identify where evidence limits the decision.
- Observe or measure the relevant feature in Cloning and .
- Organize the observation with a stable evidence ID.
- Apply this rule: Stored information can be copied, read, and converted into a functional product.
- Choose the option the evidence supports and state the limit of the conclusion.
Real biomedical example: You built on paper yesterday, but how do you force millions of bacteria to actually swallow that , and how do you find the few that did?
What the evidence supports: E1-E3 and F1 support the daily take-home when the response meets the stated success criteria.
What it cannot prove: The package does not support claims beyond this lesson's or any real patient diagnosis.
- • : A small circular piece of DNA found in bacteria that copies itself separately from the main and is often used to carry genes in the lab.
- • : DNA made by joining genetic material from two different sources, often to insert a chosen gene into a cell so it makes a useful .
- • : An that joins two pieces of DNA together by sealing the gap in their backbone, vital in genetic engineering.
- • : The process by which a bacterial cell takes up foreign DNA, such as a , from its surroundings and begins using those new genes.
- • expression: The process of turning a gene on so its DNA instructions are used to make RNA and , deciding when and where a gene is active.
Use it now: Choose one decision option. Cite E1 and E3, then explain how the rule connects the evidence to your choice.
Go further, optional: The source links below are optional enrichment. Every fact required for today's local evidence decision appears in this lesson package.
Heat shock or electroporation opens pores in bacterial membranes so DNA can enter.
Limit: E1 supplies context or an observation; it does not by itself establish the explanation.
A recombinant can be mass-produced because puts the into cells, selection keeps only the cells that took it, and separates the protein from debris, so each step sets up the next.
Limit: E2 is a teaching statement or comparison and must be checked against the task evidence.
You'll be able to carry out , selection, and steps.
Limit: E3 supports only the result or product criterion named here; it cannot justify a broader clinical or causal claim.
PLTW-GEND-2026-11-24 · Simulated classroom evidence scenario
Your role: medical interventions team member
Decision: Your team must decide what the evidence from Cloning and supports before submitting the lab report named on the lesson page.
- • Select the option best supported by E1-E3.
- • Select a reasonable alternative and name the evidence it would require.
- • Delay the claim because the evidence does not distinguish the options.
Response: State one choice, cite at least two evidence IDs, explain the rule that connects them, and add one limitation. Submit it as the lab report.
Claim ceiling: The supplied lesson evidence can support an observation, pattern, classroom mechanism, or next-step decision about Cloning and . It cannot by itself prove causation, establish a real clinical diagnosis, or justify action outside this classroom task.
Reason for review: Your team must decide what the evidence from Cloning and supports before submitting the lab report named on the lesson page.
Context: Producing a recombinant is a pipeline where each step depends on the one before: cells must take up the DNA, only the right cells are kept alive, and only then can the protein be grown and pulled out clean.
- • T1: Read the protocol in the PLTW course shell and gather your materials.
- • T2: Model by introducing the recombinant into host cells.
- • T3: Select transformed cells using the provided marker and record how many grew.
- • T4: Run the simulated step and note where the target appears.
- • T5: Record yield and one quality observation in your .
- • T6: Submit your cloning and results.
- • E1: Heat shock or electroporation opens pores in bacterial membranes so DNA can enter.
- • E2: A recombinant can be mass-produced because puts the into cells, selection keeps only the cells that took it, and separates the protein from debris, so each step sets up the next.
- • E3: You'll be able to carry out , selection, and steps.
Measurements: No patient measurement is supplied unless it appears explicitly in E1-E3 or F1. Do not invent a value.
Figure finding: Teaching diagram for Cloning and . Trace the labeled testing, treatment, or biological process and identify where evidence limits the decision. This is a teaching model, not patient or experimental data.
Uncertainty: This is a composite classroom scenario. Missing history, measurements, or confirmation tests remain unknown and limit the conclusion.
Students often think Students assume that if you add DNA to a tube of bacteria, all or most of the cells will take it up and carry the gene.. The trap: is rare; only a small fraction of cells take up the . That is exactly why selection exists: the plasmid carries a resistance gene, so the antibiotic kills the failures and only transformed cells survive. If you skip why selection is needed, the whole looks pointless.
Workflow record:
- Transformation: introduced the recombinant plasmid into host cells using heat shock, which opens pores in the membrane so plasmid DNA can enter.
- Selection: plated cells on antibiotic medium. Only cells carrying the resistance gene on the plasmid survived. I counted 38 colonies on the plate with my recombinant cells and 0 on the no-plasmid control, which tells me selection worked.
- Purification: ran the first separation step, which split soluble protein away from cell debris. The target protein appeared in the soluble fraction.
- Yield and quality: estimated yield was moderate; one quality note is that the no-plasmid control showed no growth, so the colonies I counted really are transformed cells and not contamination.
| Step | Action | Result |
|---|---|---|
| Transformation | Heat shock plasmid into cells | Cells took up plasmid |
| Selection | Plate on antibiotic | 38 colonies; control 0 |
| Purification | First separation | Target in soluble fraction |
| Quality note | Check control plate | No growth, so no contamination |
This model shows the level of evidence and organization needed to complete: Completes the workflow lab: a data table recording transformation, selection counts, fraction data, yield, and one quality observation across the cloning-to-purification pipeline.
- State the question and method.
- Present the observations and data with units.
- Explain the result, limitations, and next investigation.
Keep the structure. Replace the question, facts, measurements, and evidence. Then recheck units, vocabulary, and whether the conclusion goes beyond the evidence.
Also due today: Submit your cloning and purification workflow data table to the class site before the end of block.
- 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, name the context clue or evidence that helped, then give one example from what you actually did in Cloning and purification workflow. 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 Clever, then Schoology, and find each one by the name shown on its card.
Use this if you were absent, got stuck, or need another pass before you submit the lesson artifact.
Placement rationale
Matched and cloning by path:Medical-Interventions/Unit-4_When-Organs-Fail/4.1_Manufacturing-Human-Proteins; keywords:, pglo. Score 142. 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 cloning by path:Medical-Interventions/Unit-4_When-Organs-Fail/4.1_Manufacturing-Human-Proteins; keywords:, pglo. Score 138. 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 cloning by path:Medical-Interventions/Unit-4_When-Organs-Fail/4.1_Manufacturing-Human-Proteins; keywords:, pglo. Score 138. 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. Both myPLTW and Schoology are in Clever. Do the activity in myPLTW. Turn the work in on this site or hand it to Mr. Mendoza, because that is the step that counts as submitted. Schoology only shows your report-card grade later.
Check yourself · commit, then reveal▸
Claim ceiling for this check: The supplied lesson evidence can support an observation, pattern, classroom mechanism, or next-step decision about Cloning and . It cannot by itself prove causation, establish a real clinical diagnosis, or justify action outside this classroom task.
You spread transformed bacteria on a plate containing an antibiotic. Billions of cells were added, but only about 40 colonies grow. Why did almost all the cells die, and what do the survivors have in common?
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▸
I can name the procedure's purpose and the evidence I will record. I can identify each named hazard and the control that reduces it: Wear nitrile gloves and safety goggles throughout the procedure. My data table is ready before materials are handled.
Finish the checklist before you handle any material.
- • Wear nitrile gloves and safety goggles throughout the procedure.
- • Treat all bacterial cultures as BSL-1 organisms: avoid mouth contact and wash hands before leaving lab.
- • Dispose of all biological waste (plates, tubes, tips) in designated biohazard bags.
- • Wipe bench with 10% bleach or 70% ethanol before and after use.
- • Report any spill involving bacterial culture to the teacher immediately.
- • Do not eat, drink, or apply cosmetics in the lab area.
- 1Before materials are handled, identify the purpose, variables or comparison, controls, measurement units, and stop-work condition.
- 2Read the workflow protocol in the PLTW course shell and gather your materials.
- 3Model transformation by introducing the recombinant plasmid into host cells.
- 4Select transformed cells using the provided marker and record how many grew.
- 5Run the simulated purification step and note where the target protein appears.
- 6Record yield and one quality observation in your workflow data table.
- 7Submit your cloning and purification workflow results.
- 8Record each result in the prepared table before interpreting it. Mark missing, repeated, or invalid results truthfully.
- 9Complete the named cleanup and waste route, remove PPE safely, wash hands when required, and confirm the station is ready for the next group.
| Trial or sample ID | Independent condition | Measured result with units | Observation before interpretation | Quality-control note |
|---|---|---|---|---|
Before the procedure, predict the result and cite the rule behind the prediction.
After the procedure, compare the result with the prediction and name one limitation or source of uncertainty.
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.
Run the virtual and linked on the class site, recording selection counts and where the target elutes, then submit your workflow .
Learn.Genetics: Bacterial TransformationThen submit your Lab report. Turn this in at the drop folder with your district Microsoft sign-in, or hand it to Mr. Mendoza in class. Both count as submitted. Doing the activity in myPLTW does not.
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: CloningYou've passed Unit 2, so the optional extra-credit track is open. Complete reserved-unit work from home, including virtual labs, for extra credit. Each item shows its correct submission route.
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, on the class site or handed to Mr. Mendoza in class, and confirmed. Not in Schoology: that is where the report-card grade appears later.
- 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.