SlideShare a Scribd company logo
1 of 3
Download to read offline
Name:                                                                 Date:


                                        PGLO Lab Practice
        Cut out the descriptions of each step on the back page and match them to the pictures on the right.


1.


2.



3.




4.




5.




6.
7.


8.




9.




10.



11.



12.
Please cut out the steps below and match them with the pictures on the previous page.




A. Label one mico test tube +pGLO and the other –              B. Use a new sterile inoculating loop to spread the liquid
pGLO. The +pGLO is the one with the plasmids, the –            evenly across the surface of the agar in a zig-zag pattern.
pGLO is the control without plasmids, just to compare.         This spreads the bacteria out so it can reach all the
                                                               nutrients.

C. Heat shock. Using the foam rack as a holder, move           D. Use a sterile inoculating loop to pick up a single spot
the microtubes into the water bath at 42°C for 50              of bacteria from the Petri dish. Put the loop in the
seconds. This causes the phospholipids that make up the        microtubes and spin/twist the loop until all the bacteria
cell membrane of the bacteria to expand, allowing the          is mixed into the solution.
plasmids to enter the bacteria.

E. Place the microtubes on ice. This brings down the           F. Using a sterile pipette, add 250 microliters of LB
temperature to shrink the cell membrane of the bacteria,       Broth to each of the microtubes. This gives the bacteria
making it more stretchy during heat shock.                     extra nutrients.

G. While your tubes are sitting on ice, label your four        H. Using an new sterile pipette, add 100 microliters of
Petri dishes as +pLGO LB/amp, +pGLO LB/amp/ara,                the transformation and control suspensions onto the
-pGLO LB/amp, and –pGLO LB. Label them on the                  appropriate Petri dishes.
bottom, not the lid, so if you switch the lids accidentally,
you still know which is which.

I. Put a new sterile inoculating loop into the plasmid         J. Put the microtubes in a test tube rack, and then on ice
DNA microtube. There should be a layer of plasmid              again, for 10 minutes. This brings down the
solution across the ring. Mix it into the +pGLO                temperature to shrink the cell membrane of the bacteria,
microtube, but NOT the –pGLO microtube. We do this             making it more stretchy during heat shock.
because the –pGLO is our control, to compare and make
sure we are growing the bacteria right.

K. Using a sterile pipette, transfer 250 microliters of        L. Stack up your Petri dishes and tape them. Put them in
transformation solution (CaCl2) to the microtubes. This        an incubator at 37°C for 24 hours. This warm
gives the plasmid DNA a negative charge, which allows          temperature is the best environment for the E. coli to
it to pass through the cell membrane of the bactiera.          grow in.

More Related Content

Similar to P glo lab kinesthetic

Genetic Transformation Lab
Genetic Transformation LabGenetic Transformation Lab
Genetic Transformation LabSonja Silva
 
1 Objectives Genetically transform bacteria with for.docx
1 Objectives Genetically transform bacteria with for.docx1 Objectives Genetically transform bacteria with for.docx
1 Objectives Genetically transform bacteria with for.docxmercysuttle
 
Ampicillin Resistance Lab Report
Ampicillin Resistance Lab ReportAmpicillin Resistance Lab Report
Ampicillin Resistance Lab ReportSheila Phillips
 
The Transformation Of E. Coli
The Transformation Of E. ColiThe Transformation Of E. Coli
The Transformation Of E. ColiLana Sorrels
 
Pglo Transformation Lab Report
Pglo Transformation Lab ReportPglo Transformation Lab Report
Pglo Transformation Lab ReportApril Davis
 
Kiran b k , report of protocol for transformation of the e coli by electropor...
Kiran b k , report of protocol for transformation of the e coli by electropor...Kiran b k , report of protocol for transformation of the e coli by electropor...
Kiran b k , report of protocol for transformation of the e coli by electropor...Prajwal N R
 
Gene Expression And Its Engineering
Gene Expression And Its EngineeringGene Expression And Its Engineering
Gene Expression And Its EngineeringRenee Delgado
 
Escherichia Coli, Staphylococcus Aureus, And Enterococcus...
Escherichia Coli, Staphylococcus Aureus, And Enterococcus...Escherichia Coli, Staphylococcus Aureus, And Enterococcus...
Escherichia Coli, Staphylococcus Aureus, And Enterococcus...Sandy Grasso
 
Biol 390 – Lab 8 Restriction Digest and Gel Electrophoresis .docx
Biol 390 – Lab 8 Restriction Digest and Gel Electrophoresis .docxBiol 390 – Lab 8 Restriction Digest and Gel Electrophoresis .docx
Biol 390 – Lab 8 Restriction Digest and Gel Electrophoresis .docxmoirarandell
 
- Lab 9 Preparation for Protein PurificationObjectivesGrow tra.docx
- Lab 9 Preparation for Protein PurificationObjectivesGrow tra.docx- Lab 9 Preparation for Protein PurificationObjectivesGrow tra.docx
- Lab 9 Preparation for Protein PurificationObjectivesGrow tra.docxoswald1horne84988
 
Vernier Photosynthesis Lab #7
Vernier Photosynthesis Lab #7Vernier Photosynthesis Lab #7
Vernier Photosynthesis Lab #7Brad Kremer
 
Plasmid Transformation Lab
Plasmid Transformation LabPlasmid Transformation Lab
Plasmid Transformation LabSheila Guy
 
Lab Report On Pglo Transformation
Lab Report On Pglo TransformationLab Report On Pglo Transformation
Lab Report On Pglo TransformationBrooke Curtis
 

Similar to P glo lab kinesthetic (20)

Genetic Transformation Lab
Genetic Transformation LabGenetic Transformation Lab
Genetic Transformation Lab
 
1 Objectives Genetically transform bacteria with for.docx
1 Objectives Genetically transform bacteria with for.docx1 Objectives Genetically transform bacteria with for.docx
1 Objectives Genetically transform bacteria with for.docx
 
Lab Report Plasmid Pglo
Lab Report Plasmid PgloLab Report Plasmid Pglo
Lab Report Plasmid Pglo
 
Ampicillin Resistance Lab Report
Ampicillin Resistance Lab ReportAmpicillin Resistance Lab Report
Ampicillin Resistance Lab Report
 
The Transformation Of E. Coli
The Transformation Of E. ColiThe Transformation Of E. Coli
The Transformation Of E. Coli
 
Pcr Lab Report
Pcr Lab ReportPcr Lab Report
Pcr Lab Report
 
Pglo Transformation Lab Report
Pglo Transformation Lab ReportPglo Transformation Lab Report
Pglo Transformation Lab Report
 
Kiran b k , report of protocol for transformation of the e coli by electropor...
Kiran b k , report of protocol for transformation of the e coli by electropor...Kiran b k , report of protocol for transformation of the e coli by electropor...
Kiran b k , report of protocol for transformation of the e coli by electropor...
 
GFP Write-up
GFP Write-upGFP Write-up
GFP Write-up
 
Gene Expression And Its Engineering
Gene Expression And Its EngineeringGene Expression And Its Engineering
Gene Expression And Its Engineering
 
Escherichia Coli, Staphylococcus Aureus, And Enterococcus...
Escherichia Coli, Staphylococcus Aureus, And Enterococcus...Escherichia Coli, Staphylococcus Aureus, And Enterococcus...
Escherichia Coli, Staphylococcus Aureus, And Enterococcus...
 
Biol 390 – Lab 8 Restriction Digest and Gel Electrophoresis .docx
Biol 390 – Lab 8 Restriction Digest and Gel Electrophoresis .docxBiol 390 – Lab 8 Restriction Digest and Gel Electrophoresis .docx
Biol 390 – Lab 8 Restriction Digest and Gel Electrophoresis .docx
 
PGLO expirement
PGLO expirementPGLO expirement
PGLO expirement
 
- Lab 9 Preparation for Protein PurificationObjectivesGrow tra.docx
- Lab 9 Preparation for Protein PurificationObjectivesGrow tra.docx- Lab 9 Preparation for Protein PurificationObjectivesGrow tra.docx
- Lab 9 Preparation for Protein PurificationObjectivesGrow tra.docx
 
Plasmid Lab Report
Plasmid Lab ReportPlasmid Lab Report
Plasmid Lab Report
 
Bacterial Lawns
Bacterial LawnsBacterial Lawns
Bacterial Lawns
 
Vernier Photosynthesis Lab #7
Vernier Photosynthesis Lab #7Vernier Photosynthesis Lab #7
Vernier Photosynthesis Lab #7
 
Plasmid Transformation Lab
Plasmid Transformation LabPlasmid Transformation Lab
Plasmid Transformation Lab
 
Lab Report On Pglo Transformation
Lab Report On Pglo TransformationLab Report On Pglo Transformation
Lab Report On Pglo Transformation
 
Pglo Plasmid Experiment
Pglo Plasmid ExperimentPglo Plasmid Experiment
Pglo Plasmid Experiment
 

More from Mills Cbst

P glo worksheet
P glo worksheetP glo worksheet
P glo worksheetMills Cbst
 
Match ups answers
Match ups answersMatch ups answers
Match ups answersMills Cbst
 
Manatee fact sheet
Manatee fact sheetManatee fact sheet
Manatee fact sheetMills Cbst
 
Elephant evolution tree
Elephant evolution treeElephant evolution tree
Elephant evolution treeMills Cbst
 
Dugong fact sheet
Dugong fact sheetDugong fact sheet
Dugong fact sheetMills Cbst
 
Evolution beaks modified worksheet
Evolution beaks modified worksheetEvolution beaks modified worksheet
Evolution beaks modified worksheetMills Cbst
 
Bird beaks lesson
Bird beaks lessonBird beaks lesson
Bird beaks lessonMills Cbst
 
pGLO tutorial en español
pGLO tutorial en español  pGLO tutorial en español
pGLO tutorial en español Mills Cbst
 
pGLO Tutorial - English
pGLO Tutorial - English pGLO Tutorial - English
pGLO Tutorial - English Mills Cbst
 
Barrett rosa meza-acevedo & lingou
Barrett rosa meza-acevedo & lingouBarrett rosa meza-acevedo & lingou
Barrett rosa meza-acevedo & lingouMills Cbst
 
Barrett presentation valerie metea
Barrett presentation valerie meteaBarrett presentation valerie metea
Barrett presentation valerie meteaMills Cbst
 
Stefanie yoshizuka pzazz
Stefanie yoshizuka pzazzStefanie yoshizuka pzazz
Stefanie yoshizuka pzazzMills Cbst
 
Monica's Barrett Presentation
Monica's Barrett PresentationMonica's Barrett Presentation
Monica's Barrett PresentationMills Cbst
 
Mo's Barret Presentation
Mo's Barret PresentationMo's Barret Presentation
Mo's Barret PresentationMills Cbst
 
Congreso de Biotecnología Arequipa Perú June 2011
Congreso de Biotecnología Arequipa Perú June 2011Congreso de Biotecnología Arequipa Perú June 2011
Congreso de Biotecnología Arequipa Perú June 2011Mills Cbst
 
TTT conference presentation ENGLISH
TTT conference presentation ENGLISHTTT conference presentation ENGLISH
TTT conference presentation ENGLISHMills Cbst
 

More from Mills Cbst (18)

P glo worksheet
P glo worksheetP glo worksheet
P glo worksheet
 
Match ups answers
Match ups answersMatch ups answers
Match ups answers
 
Manatee fact sheet
Manatee fact sheetManatee fact sheet
Manatee fact sheet
 
Match ups
Match upsMatch ups
Match ups
 
Elephant evolution tree
Elephant evolution treeElephant evolution tree
Elephant evolution tree
 
Dugong fact sheet
Dugong fact sheetDugong fact sheet
Dugong fact sheet
 
Evolution beaks modified worksheet
Evolution beaks modified worksheetEvolution beaks modified worksheet
Evolution beaks modified worksheet
 
Bird beaks lesson
Bird beaks lessonBird beaks lesson
Bird beaks lesson
 
pGLO tutorial en español
pGLO tutorial en español  pGLO tutorial en español
pGLO tutorial en español
 
pGLO Tutorial - English
pGLO Tutorial - English pGLO Tutorial - English
pGLO Tutorial - English
 
Barrett rosa meza-acevedo & lingou
Barrett rosa meza-acevedo & lingouBarrett rosa meza-acevedo & lingou
Barrett rosa meza-acevedo & lingou
 
Barrett presentation valerie metea
Barrett presentation valerie meteaBarrett presentation valerie metea
Barrett presentation valerie metea
 
Stefanie yoshizuka pzazz
Stefanie yoshizuka pzazzStefanie yoshizuka pzazz
Stefanie yoshizuka pzazz
 
Monica's Barrett Presentation
Monica's Barrett PresentationMonica's Barrett Presentation
Monica's Barrett Presentation
 
Mo's Barret Presentation
Mo's Barret PresentationMo's Barret Presentation
Mo's Barret Presentation
 
Congreso de Biotecnología Arequipa Perú June 2011
Congreso de Biotecnología Arequipa Perú June 2011Congreso de Biotecnología Arequipa Perú June 2011
Congreso de Biotecnología Arequipa Perú June 2011
 
NSF highlight
NSF highlightNSF highlight
NSF highlight
 
TTT conference presentation ENGLISH
TTT conference presentation ENGLISHTTT conference presentation ENGLISH
TTT conference presentation ENGLISH
 

Recently uploaded

Comparing Sidecar-less Service Mesh from Cilium and Istio
Comparing Sidecar-less Service Mesh from Cilium and IstioComparing Sidecar-less Service Mesh from Cilium and Istio
Comparing Sidecar-less Service Mesh from Cilium and IstioChristian Posta
 
UWB Technology for Enhanced Indoor and Outdoor Positioning in Physiological M...
UWB Technology for Enhanced Indoor and Outdoor Positioning in Physiological M...UWB Technology for Enhanced Indoor and Outdoor Positioning in Physiological M...
UWB Technology for Enhanced Indoor and Outdoor Positioning in Physiological M...UbiTrack UK
 
Cybersecurity Workshop #1.pptx
Cybersecurity Workshop #1.pptxCybersecurity Workshop #1.pptx
Cybersecurity Workshop #1.pptxGDSC PJATK
 
IaC & GitOps in a Nutshell - a FridayInANuthshell Episode.pdf
IaC & GitOps in a Nutshell - a FridayInANuthshell Episode.pdfIaC & GitOps in a Nutshell - a FridayInANuthshell Episode.pdf
IaC & GitOps in a Nutshell - a FridayInANuthshell Episode.pdfDaniel Santiago Silva Capera
 
AI You Can Trust - Ensuring Success with Data Integrity Webinar
AI You Can Trust - Ensuring Success with Data Integrity WebinarAI You Can Trust - Ensuring Success with Data Integrity Webinar
AI You Can Trust - Ensuring Success with Data Integrity WebinarPrecisely
 
Building AI-Driven Apps Using Semantic Kernel.pptx
Building AI-Driven Apps Using Semantic Kernel.pptxBuilding AI-Driven Apps Using Semantic Kernel.pptx
Building AI-Driven Apps Using Semantic Kernel.pptxUdaiappa Ramachandran
 
Designing A Time bound resource download URL
Designing A Time bound resource download URLDesigning A Time bound resource download URL
Designing A Time bound resource download URLRuncy Oommen
 
9 Steps For Building Winning Founding Team
9 Steps For Building Winning Founding Team9 Steps For Building Winning Founding Team
9 Steps For Building Winning Founding TeamAdam Moalla
 
Linked Data in Production: Moving Beyond Ontologies
Linked Data in Production: Moving Beyond OntologiesLinked Data in Production: Moving Beyond Ontologies
Linked Data in Production: Moving Beyond OntologiesDavid Newbury
 
COMPUTER 10: Lesson 7 - File Storage and Online Collaboration
COMPUTER 10: Lesson 7 - File Storage and Online CollaborationCOMPUTER 10: Lesson 7 - File Storage and Online Collaboration
COMPUTER 10: Lesson 7 - File Storage and Online Collaborationbruanjhuli
 
20230202 - Introduction to tis-py
20230202 - Introduction to tis-py20230202 - Introduction to tis-py
20230202 - Introduction to tis-pyJamie (Taka) Wang
 
UiPath Platform: The Backend Engine Powering Your Automation - Session 1
UiPath Platform: The Backend Engine Powering Your Automation - Session 1UiPath Platform: The Backend Engine Powering Your Automation - Session 1
UiPath Platform: The Backend Engine Powering Your Automation - Session 1DianaGray10
 
UiPath Studio Web workshop series - Day 7
UiPath Studio Web workshop series - Day 7UiPath Studio Web workshop series - Day 7
UiPath Studio Web workshop series - Day 7DianaGray10
 
Nanopower In Semiconductor Industry.pdf
Nanopower  In Semiconductor Industry.pdfNanopower  In Semiconductor Industry.pdf
Nanopower In Semiconductor Industry.pdfPedro Manuel
 
UiPath Studio Web workshop series - Day 8
UiPath Studio Web workshop series - Day 8UiPath Studio Web workshop series - Day 8
UiPath Studio Web workshop series - Day 8DianaGray10
 
ADOPTING WEB 3 FOR YOUR BUSINESS: A STEP-BY-STEP GUIDE
ADOPTING WEB 3 FOR YOUR BUSINESS: A STEP-BY-STEP GUIDEADOPTING WEB 3 FOR YOUR BUSINESS: A STEP-BY-STEP GUIDE
ADOPTING WEB 3 FOR YOUR BUSINESS: A STEP-BY-STEP GUIDELiveplex
 
How Accurate are Carbon Emissions Projections?
How Accurate are Carbon Emissions Projections?How Accurate are Carbon Emissions Projections?
How Accurate are Carbon Emissions Projections?IES VE
 
KubeConEU24-Monitoring Kubernetes and Cloud Spend with OpenCost
KubeConEU24-Monitoring Kubernetes and Cloud Spend with OpenCostKubeConEU24-Monitoring Kubernetes and Cloud Spend with OpenCost
KubeConEU24-Monitoring Kubernetes and Cloud Spend with OpenCostMatt Ray
 
UiPath Studio Web workshop series - Day 6
UiPath Studio Web workshop series - Day 6UiPath Studio Web workshop series - Day 6
UiPath Studio Web workshop series - Day 6DianaGray10
 

Recently uploaded (20)

Comparing Sidecar-less Service Mesh from Cilium and Istio
Comparing Sidecar-less Service Mesh from Cilium and IstioComparing Sidecar-less Service Mesh from Cilium and Istio
Comparing Sidecar-less Service Mesh from Cilium and Istio
 
UWB Technology for Enhanced Indoor and Outdoor Positioning in Physiological M...
UWB Technology for Enhanced Indoor and Outdoor Positioning in Physiological M...UWB Technology for Enhanced Indoor and Outdoor Positioning in Physiological M...
UWB Technology for Enhanced Indoor and Outdoor Positioning in Physiological M...
 
Cybersecurity Workshop #1.pptx
Cybersecurity Workshop #1.pptxCybersecurity Workshop #1.pptx
Cybersecurity Workshop #1.pptx
 
IaC & GitOps in a Nutshell - a FridayInANuthshell Episode.pdf
IaC & GitOps in a Nutshell - a FridayInANuthshell Episode.pdfIaC & GitOps in a Nutshell - a FridayInANuthshell Episode.pdf
IaC & GitOps in a Nutshell - a FridayInANuthshell Episode.pdf
 
20150722 - AGV
20150722 - AGV20150722 - AGV
20150722 - AGV
 
AI You Can Trust - Ensuring Success with Data Integrity Webinar
AI You Can Trust - Ensuring Success with Data Integrity WebinarAI You Can Trust - Ensuring Success with Data Integrity Webinar
AI You Can Trust - Ensuring Success with Data Integrity Webinar
 
Building AI-Driven Apps Using Semantic Kernel.pptx
Building AI-Driven Apps Using Semantic Kernel.pptxBuilding AI-Driven Apps Using Semantic Kernel.pptx
Building AI-Driven Apps Using Semantic Kernel.pptx
 
Designing A Time bound resource download URL
Designing A Time bound resource download URLDesigning A Time bound resource download URL
Designing A Time bound resource download URL
 
9 Steps For Building Winning Founding Team
9 Steps For Building Winning Founding Team9 Steps For Building Winning Founding Team
9 Steps For Building Winning Founding Team
 
Linked Data in Production: Moving Beyond Ontologies
Linked Data in Production: Moving Beyond OntologiesLinked Data in Production: Moving Beyond Ontologies
Linked Data in Production: Moving Beyond Ontologies
 
COMPUTER 10: Lesson 7 - File Storage and Online Collaboration
COMPUTER 10: Lesson 7 - File Storage and Online CollaborationCOMPUTER 10: Lesson 7 - File Storage and Online Collaboration
COMPUTER 10: Lesson 7 - File Storage and Online Collaboration
 
20230202 - Introduction to tis-py
20230202 - Introduction to tis-py20230202 - Introduction to tis-py
20230202 - Introduction to tis-py
 
UiPath Platform: The Backend Engine Powering Your Automation - Session 1
UiPath Platform: The Backend Engine Powering Your Automation - Session 1UiPath Platform: The Backend Engine Powering Your Automation - Session 1
UiPath Platform: The Backend Engine Powering Your Automation - Session 1
 
UiPath Studio Web workshop series - Day 7
UiPath Studio Web workshop series - Day 7UiPath Studio Web workshop series - Day 7
UiPath Studio Web workshop series - Day 7
 
Nanopower In Semiconductor Industry.pdf
Nanopower  In Semiconductor Industry.pdfNanopower  In Semiconductor Industry.pdf
Nanopower In Semiconductor Industry.pdf
 
UiPath Studio Web workshop series - Day 8
UiPath Studio Web workshop series - Day 8UiPath Studio Web workshop series - Day 8
UiPath Studio Web workshop series - Day 8
 
ADOPTING WEB 3 FOR YOUR BUSINESS: A STEP-BY-STEP GUIDE
ADOPTING WEB 3 FOR YOUR BUSINESS: A STEP-BY-STEP GUIDEADOPTING WEB 3 FOR YOUR BUSINESS: A STEP-BY-STEP GUIDE
ADOPTING WEB 3 FOR YOUR BUSINESS: A STEP-BY-STEP GUIDE
 
How Accurate are Carbon Emissions Projections?
How Accurate are Carbon Emissions Projections?How Accurate are Carbon Emissions Projections?
How Accurate are Carbon Emissions Projections?
 
KubeConEU24-Monitoring Kubernetes and Cloud Spend with OpenCost
KubeConEU24-Monitoring Kubernetes and Cloud Spend with OpenCostKubeConEU24-Monitoring Kubernetes and Cloud Spend with OpenCost
KubeConEU24-Monitoring Kubernetes and Cloud Spend with OpenCost
 
UiPath Studio Web workshop series - Day 6
UiPath Studio Web workshop series - Day 6UiPath Studio Web workshop series - Day 6
UiPath Studio Web workshop series - Day 6
 

P glo lab kinesthetic

  • 1. Name: Date: PGLO Lab Practice Cut out the descriptions of each step on the back page and match them to the pictures on the right. 1. 2. 3. 4. 5. 6.
  • 3. Please cut out the steps below and match them with the pictures on the previous page. A. Label one mico test tube +pGLO and the other – B. Use a new sterile inoculating loop to spread the liquid pGLO. The +pGLO is the one with the plasmids, the – evenly across the surface of the agar in a zig-zag pattern. pGLO is the control without plasmids, just to compare. This spreads the bacteria out so it can reach all the nutrients. C. Heat shock. Using the foam rack as a holder, move D. Use a sterile inoculating loop to pick up a single spot the microtubes into the water bath at 42°C for 50 of bacteria from the Petri dish. Put the loop in the seconds. This causes the phospholipids that make up the microtubes and spin/twist the loop until all the bacteria cell membrane of the bacteria to expand, allowing the is mixed into the solution. plasmids to enter the bacteria. E. Place the microtubes on ice. This brings down the F. Using a sterile pipette, add 250 microliters of LB temperature to shrink the cell membrane of the bacteria, Broth to each of the microtubes. This gives the bacteria making it more stretchy during heat shock. extra nutrients. G. While your tubes are sitting on ice, label your four H. Using an new sterile pipette, add 100 microliters of Petri dishes as +pLGO LB/amp, +pGLO LB/amp/ara, the transformation and control suspensions onto the -pGLO LB/amp, and –pGLO LB. Label them on the appropriate Petri dishes. bottom, not the lid, so if you switch the lids accidentally, you still know which is which. I. Put a new sterile inoculating loop into the plasmid J. Put the microtubes in a test tube rack, and then on ice DNA microtube. There should be a layer of plasmid again, for 10 minutes. This brings down the solution across the ring. Mix it into the +pGLO temperature to shrink the cell membrane of the bacteria, microtube, but NOT the –pGLO microtube. We do this making it more stretchy during heat shock. because the –pGLO is our control, to compare and make sure we are growing the bacteria right. K. Using a sterile pipette, transfer 250 microliters of L. Stack up your Petri dishes and tape them. Put them in transformation solution (CaCl2) to the microtubes. This an incubator at 37°C for 24 hours. This warm gives the plasmid DNA a negative charge, which allows temperature is the best environment for the E. coli to it to pass through the cell membrane of the bactiera. grow in.