Transformation and gel wet lab
Open your materials, follow the steps, then turn in your work.
Run a bacterial transformation and a gel electrophoresis to separate DNA fragments.
1. Open your materials
Use the materials named in the first step below. Open lesson resources.
2. Start the work
Follow safety protocol and prepare your competent cells and plasmid.
Show all 5 required steps
- Follow safety protocol and prepare your competent cells and plasmid.
- Perform the heat-shock transformation and plate on selective and control media.
- Set up a restriction digest of plasmid DNA.
- Load digested samples and a DNA ladder into the agarose gel.
- Run the gel and record band positions.
Lost your place? If class got interrupted, check whether your plates are already in the incubator. If yes, move to 'load digested samples and a DNA ladder into the agarose gel'; if no, restart at the heat-shock transformation step with fresh competent cells.
Check your work before submitting
- You can run a transformation with its positive and negative controls.
- Your gel separated fragments alongside a DNA ladder.
Before lab work: read the safety rules
- Wear gloves, goggles, and lab coat for the entire lab; remove before leaving the lab area.
- All bacterial cultures and plates are biohazardous: do not bring outside the lab, do not touch your face.
- Decontaminate all materials that contact bacteria with 10 percent bleach for at least 20 minutes before disposal.
- Autoclave or bleach-treat all transformed plates and liquid cultures; do not put in regular trash.
- Use UV transilluminator only with proper eye protection; UV causes corneal and skin damage.
- If using ethidium bromide, handle only in designated area; it is a mutagen and requires dedicated waste disposal.
- Report any spill, broken glass, or skin contact with reagents to the teacher immediately.
- Wash hands thoroughly with soap and water before leaving the lab.
- The water bath is hot: use tube holders, not bare hands, when removing samples.
- Keep gel apparatus lid closed while power is on; electrical shock risk from buffer contact.
3. Turn in your work
DueCheck Schoology- Hand in
- Wet lab data record: transformation plate colony counts for each condition, gel photograph with labeled lanes, raw band position measurements, and one result comparison to pre-lab prediction.
How to submit and name your file
Use the submission route shown on today's today's page.
In Schoology, open your course and the assignment for this lesson. Attach your file, select Submit, and check that it appears in the submission.
PDF upload helpYou get two school days for every day you were absent, so this deadline moves with you.
How this lesson connects
Keep using what you learned last class: Biotechnology needs institutional oversight because even small-scale work spreads real biosafety risk (like antibiotic-resistance genes) when containment fails, so the rule must follow the organism rather than the size of the lab. Today: Transformation and gel electrophoresis are the foundational recombinant-DNA techniques because one inserts new DNA into cells and the other separates DNA by size, so together they let you build a construct and then verify it.
Optional: listen or watch a unit review▸
Need help? Warm-up, timing, and directions▸
💡 Big idea: and are the foundational recombinant-DNA techniques because one inserts new DNA into cells and the other separates DNA by size, so together they let you build a construct and then verify it.
- 0-10 min briefing; confirm PPE; review and gel procedures
- 10-30 minPrepare competent cells and ; perform heat-shock ; plate on selective and control media
- 30-50 minSet up restriction digest; prepare gel; load samples and
- 50-65 minRun gel; photograph result; record band positions in lab notebook
- 65-75 minInitiate cleanup; or bleach-treat all transformed cultures per protocol
- 75-80 minExit ticket: note one result that matched and one that surprised you
- • This is a wet lab day. Everyone follows the protocol before touching any reagents.
- • We run two procedures back to back: heat-shock and .
- • Your pre-lab predictions from yesterday are your roadmap: check them against your real results.
- • Record everything as you go; data recorded after the fact is not reliable.
- • separates DNA fragments by size: smaller fragments migrate farther through the gel matrix.
- • A provides known fragment sizes that allow you to estimate unknown band sizes by comparison.
- • Plate controls verify both that the worked and that is absent.
PLTW connection and today's work
Open Problem 6 in your myPLTW course shell and navigate to the transformation and gel activity, then run the wet lab and record raw plate data and gel band positions.
Today's stopping point: The transformation notes are done; wet lab data collection is the central Problem 6 milestone, so check your activity guide.
PLTW activity titles identify the course connection. If your account will not open, use the posted materials for today and tell Mr. Mendoza. Do not mark an online activity complete unless you completed it.
Course connection
- Project 6.1.2 Construction and Cloning of Recombinant DNA
Use the turn-in directions at the top of this page. Do not create a second submission unless your teacher asks for one.
Show another explanation or a smaller first step
Need help? Choose a starting point
Finish the assigned lab safely before starting extra practice.
Lesson resources: reading, slides, 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.
Biotechnology needs institutional oversight because even small-scale work spreads real biosafety risk (like -resistance genes) when containment fails, so the rule must follow the organism rather than the size of the lab.
and are the foundational recombinant-DNA techniques because one inserts new DNA into cells and the other separates DNA by size, so together they let you build a construct and then verify it.
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: Today you run a heat-shock and cut a with restriction enzymes, then load the pieces on a gel. Which plate controls prove your transformation and your sterile technique both worked, and why do the smaller DNA fragments end up farthest down the gel?
What you already know: Biotechnology needs institutional oversight because even small-scale work spreads real biosafety risk (like -resistance genes) when containment fails, so the rule must follow the organism rather than the size of the lab.
New idea: and are the foundational recombinant-DNA techniques because one inserts new DNA into cells and the other separates DNA by size, so together they let you build a construct and then verify it.
Visual or model: F1. F1. A lesson illustration or teaching diagram for Transformation and gel wet lab. Use it with E1-E3; it is a model or context image, not experimental or patient data. What to notice: Trace the labeled system, test, or design relationship and identify which evidence should trigger revision.
- Observe or measure the relevant feature in today's lesson.
- 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: Today you run a heat-shock and cut a with restriction enzymes, then load the pieces on a gel. Which plate controls prove your transformation and your sterile technique both worked, and why do the smaller DNA fragments end up farthest down the gel?
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.
- • : The process by which a bacterial cell takes up foreign DNA, such as a , from its surroundings and begins using those new genes.
- • selection: The process of choosing among options using set criteria, such as picking the best design solution or the strongest candidate.
- • : A visible cluster of identical microorganisms growing on a plate, all descended from a single original cell.
- • digest: To cut DNA at specific sequences using restriction enzymes, producing defined fragments that scientists can sort, study, or join together.
- • : A lab method that uses an electric field to pull DNA fragments through a gel so they separate by size and show up as bands.
- • : A set of DNA fragments of known sizes run alongside samples in to measure the length of unknown bands.
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.
Molecular workflows use controls and complementary separation or detection steps to evaluate a target, while each band, fraction, or remains limited by method resolution and sample quality.
Limit: A band, selected , or fluorescent fraction alone does not prove identity, , correct sequence, biological activity, or clinical suitability.
Stored information can be copied, read, and converted into a functional product.
Limit: Genes are regulated biological sequences, not conscious instructions, and one gene rarely determines a whole trait alone.
You can run a with its positive and negative controls.
Limit: E3 defines the classroom product or success criterion. It is not independent scientific evidence and cannot justify a clinical or causal claim.
PLTW-BFH-2027-05-06 · Simulated classroom evidence scenario
Your role: biomedical design team member
Decision: Your team must decide what the evidence from today's lesson supports before submitting the lab report named on today's page.
- • Read the farthest bands as your largest fragments, because heavier DNA pushes harder and travels farther through the gel.
- • A on the selection plate shows entered, not that the right insert came with it, so confirmation is missing.
- • Expect the smallest fragments farthest down the gel, since the mesh slows big pieces and lets small ones through.
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: Today's evidence supports a classroom claim about today's lesson. It cannot prove causation, diagnose a real patient, or justify action outside this room.
Rate or percent = part / comparison total x 100%. Percent change = (new - comparison) / comparison x 100%.
If 18 of 60 records meet a condition, the frequency is 18 / 60 x 100% = 30%.
Name the comparison total. A percent describes the supplied group and does not automatically predict an individual's outcome.
Use today's supplied counts to calculate one rate, risk, frequency, or percent change. Show the denominator and interpretation.
- • The solution must address the stated need in today's lesson.
- • The decision must be supported by E1-E3.
- • The final product must make the success criteria visible.
- • Complete the work inside the 80-minute block.
- • Use only supplied or teacher-approved materials and evidence.
- • Do not trade , accessibility, or privacy for speed.
- • and evidence quality: must pass before scoring other criteria.
- • User need and effectiveness: highest scored criterion.
- • Time, cost, and ease of use: compare only after and effectiveness pass.
Test evidence: For each option, record the E1-E3 result that supports or fails each criterion. Do not assign a score without a named observation.
- Version or option tested
- Criterion met or missed
- Evidence ID and result
- Revision made
- Reason for the revision
- Need and user
- Criteria and constraints
- Chosen option and evidence
- Test result
- Revision and reason
Students often think Students expect larger DNA fragments to move farther 'because they are heavier and push through harder,' picturing it like dropping rocks.. The trap: It is the opposite. The gel is a dense mesh, so large fragments snag and lag while small fragments thread through and race ahead. distance is inversely related to size, because the matrix filters by how easily a fragment squeezes through, not by weight.
Transformation results (colony counts):
- Cells + plasmid, antibiotic agar: 84 colonies
- Cells + plasmid, plain agar: lawn (too many to count)
- Cells, no plasmid, antibiotic agar: 0 colonies
- Cells, no plasmid, plain agar: lawn
Gel setup: Lane 1 = DNA ladder, Lane 2 = uncut plasmid, Lane 3 = plasmid cut with one enzyme, Lane 4 = plasmid cut with two enzymes.
Raw band measurements (distance migrated from well):
- Lane 3: one band at 22 mm
- Lane 4: bands at 18 mm and 31 mm
Comparison to prediction: My pre-lab predicted no growth on the no-plasmid antibiotic plate, and I observed 0 colonies, which matches. The single-cut lane showing one band also matches the prediction that cutting a circular plasmid once gives one linear fragment.
| Condition | Plasmid | Antibiotic | Colony count |
|---|---|---|---|
| A | Yes | Yes | 84 |
| B | Yes | No | Lawn |
| C | No | Yes | 0 |
| D | No | No | Lawn |
This model shows the level of evidence and organization needed to complete: Completes the wet-lab data page: transformation plate colony counts for each condition, a gel photo with labeled lanes, raw band-position measurements, and one comparison to the pre-lab prediction.
- 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 lab notebook data page on Schoology by end of class.
- 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 Transformation and gel wet 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 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 , , molecular evidence by path:Biomedical-Innovations/Problem-6_Molecular-Biology/6.1_Molecular-Biology; keywords:transformation, , molecular. Score 146. 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 , , molecular evidence by path:Biomedical-Innovations/Problem-6_Molecular-Biology/6.1_Molecular-Biology; keywords:gel, molecular. 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 , , molecular evidence by path:Biomedical-Innovations/Problem-6_Molecular-Biology/6.1_Molecular-Biology; keywords:transformation, molecular. Score 142. Visibility: student-schoology (student-facing resource; link through Schoology rather than local path).
Sign in to Clever with your district Microsoft account to open Schoology or myPLTW. Follow today's posted steps. If myPLTW will not open, use the posted alternative and tell Mr. Mendoza. Turn in your completed work through the Schoology assignment.
Practice: try a question, then check your answer▸
Claim ceiling for this check: Today's evidence supports a classroom claim about today's lesson. It cannot prove causation, diagnose a real patient, or justify action outside this room.
Your gel shows three sample bands. The one nearest the wells and the one nearest the bottom edge are both DNA fragments. Which fragment is larger, and how do you know?
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.
Missed class or ready for more?▸
Run this before you touch the bench. It is built from the real lab procedure, so the decisions you make here are the ones you will make with the equipment in your hands.
I can name the procedure's purpose and the evidence I will record. I can name today's hazards and the control for each: Report any spill, broken glass, or skin contact with reagents to the teacher immediately. My data table is ready before materials are handled.
Finish the checklist before you handle any material.
- • Wear gloves, goggles, and lab coat for the entire lab; remove before leaving the lab area.
- • All bacterial cultures and plates are biohazardous: do not bring outside the lab, do not touch your face.
- • Decontaminate all materials that contact bacteria with 10 percent bleach for at least 20 minutes before disposal.
- • Autoclave or bleach-treat all transformed plates and liquid cultures; do not put in regular trash.
- • Use UV transilluminator only with proper eye protection; UV causes corneal and skin damage.
- • If using ethidium bromide, handle only in designated area; it is a mutagen and requires dedicated waste disposal.
- • Report any spill, broken glass, or skin contact with reagents to the teacher immediately.
- • Wash hands thoroughly with soap and water before leaving the lab.
- • The water bath is hot: use tube holders, not bare hands, when removing samples.
- • Keep gel apparatus lid closed while power is on; electrical shock risk from buffer contact.
- 1Before materials are handled, identify the purpose, variables or comparison, controls, measurement units, and stop-work condition.
- 2Follow safety protocol and prepare your competent cells and plasmid.
- 3Perform the heat-shock transformation and plate on selective and control media.
- 4Set up a restriction digest of plasmid DNA.
- 5Load digested samples and a DNA ladder into the agarose gel.
- 6Run the gel and record band positions.
- 7Record each result in the prepared table before interpreting it. Mark missing, repeated, or invalid results truthfully.
- 8Complete 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.
Analyze the provided plate photos and gel images: count colonies on each plate and estimate fragment sizes against the ladder.
Learn.Genetics gel electrophoresisUse the submission route shown on today's today's page.
Class still runs. Complete the online activity above (it's self-guided). Need the concept taught without a teacher? Use this authoritative explainer:
Learn.Genetics (University of Utah): gel electrophoresisYou'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.
- SubmittedGo to Schoology to turn this in. Submit one PDF. Put your first and last name in the document header. Name the file: FirstName LastName - Assignment Title - YYYY-MM-DD.pdf. If you cannot get in, see Mr. Mendoza. Do not skip the work.
- 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.
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