Use the loop and streak cells on the first line, from 1A-1B.
Unlock the next clean loop by clicking on the discard bin
1. Make LB Agar Plates
Congrats! You successfully streaked K12 E. coli cells onto your non-selective LB agar plates! Its time to incubate them
One more thing…. You have to flip your plate so the lid is down. This makes sure that moisture floats up and keeps the surface of the LB agar moist. Cells like to be moist!
Flip the plate
Ok, now incubate
Success! You’ve grown “blank” cells. We call them “blank” because they are not engineered yet. Next, you’ll pick some of these cells and mix them with a special buffer to make what we call “competent” cells. Because of the special buffer, these cells will be able take in DNA during the transformation.
Next Step: Competent Cells
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The DNA Playground has temperature and incubation capabilities that you need to engineer and grow cells!
You'll use the Blank Cells you grew. Well separated colonies grow fast and fast growing cells take up DNA best!
Blue inoculating loops are sterile loops, similar to the yellow one you used before. The only difference is the loop is smaller.
Transformation buffer is water mixed with some special salts. One of the main salts is calcium chloride (CaCl2). The positive calcium ions can bind to the negatively charged DNA to help it enter cells.
You’re going to genetically engineer cells using a procedure called a “colony transformation”. This is when you collect “Blank Cell" colonies and then “transform” them with a DNA plasmid.
Start by turning on your Cold Station so that it is cold when you need it! “Ice cold” environments are frequently used in science research to keep samples stable and protect them from degrading
Place your Transformation Buffer on the Cold Station so that it is cold when you add the Blank Cells. It is very important that the Cells & Transformation Buffer are cold throughout the experiment! This keeps them stable.
Collect Blank Cells
While your Transformation Buffer is cooling down, you can collect some Blank Cells.
The Blank cells that you’ll engineer must be growing fast. Individual colonies grow the fastest. You’ll need about 10 small colonies. 10 colonies will amount to millions of bacteria! This is why streaking your plate is an important step!
Start Collecting Cells
A good amount of cells to collect is to fill the centre of the blue 1 uL loop with cells.
You have to put the cells into Transformation Buffer that is COLD. If you put the cells into warm buffer, they’ll die.
Dislodge the cells from the loop by twisting the inoculating loop vigorously. Keep the tube cold in the Cold Station! Vigorously twist so that it makes the solution cloudy and there are no clumps of cells visible.
Twist the Loop
If there are cellular clumps, then the cells inside the clumps cannot interact with the transformation buffer nor the DNA you’ll add in the following step! If this happens, they cannot be transformed! Mixing well is a very important part of doing scienice experiments
Make sure the cells are stay on the Cold Station!
Your cells are now able to take in DNA. You should move onto the Transformation step immediately! If you wait more than a two minutes before adding your DNA, your transformation efficiency declines.
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Next Step: Transform Cells
You also want to mix the cells enough so they fully mix and are suspended (not in clumps).
The DNA Playground has cold and warm stations used for heat shocking during a Transformation, as well as an incubator for incubating your Blank and Engineered Cells.
See DNA Program
Thank you, Friend! Got it
Hi Friend, All living things have DNA that informs the cells how to grow, what to produce, and how to interact with other cells and the environment. You can use DNA to engineer cells. In this case you’ll use a circular DNA molecule called a plasmid. Your DNA will “program" the cells to make a colourful pigment that originally comes from coral in the ocean.
I’ve made this DNA plasmid for you so that you can engineer E. coli cells to produce your coral pigment and turn Red. If you succeed, we may be able to grow vast amounts of your bacteria and make vast amounts of red ink.
You’ll use a blue inoculation loop to transfer DNA into the tube of competent cells. The blue loops can hold 1 uL of liquid.
Your chemically competent cells (cells that can take up DNA) should still be cold. You’ll insert a DNA program (plasmid) into them to change their genetic makeup, and result in the “expression” of a trait
Your Cold Station is still on, keeping your Competent Cells cold. You need to also turn on the 42 ℃ Hot Station for the Heat Shock step of the Transformation. A heat shock is when you change the temperature of the cells from cold to warm. This helps to cause your DNA plasmid to enter the cells.
Use the blue inoculating loop to get 1 uL of liquid containing DNA by simply dipping and twisting it. A micropipet can also be used, but because this is our first genetic engineering experiment, we’ll use a loop.
You’ll need to inspect the loop to make sure that DNA got into the loop.
Inspect the Loop
When you take your loop out of the DNA tube, check to see if there is liquid in the loop. The blue loops can hold 1 uL of liquid.
When doing science research, you should repeatedly double check your actions to make sure you are doing them properly.
Use the blue inoculating loop to get 1 uL of DNA, by simply dipping it into the DNA
Dip the loop with DNA into your Competent Cells while they remain on the Cold Station. Spin it to mix the DNA with the cells. Again, mixing is an important part of good experimentation. This will cause the DNA plasmid to enter the liquid and bump around.
Spin the Loop
When you are done spinning and mixing in the DNA for 10 seconds, discard the loop.
Incubate 5 Mintues
Incubate the Competent Cells & DNA on the Cold Station for 5 minutes. During this time the DNA binds to the cells with help from the salts inside the buffer. Some DNA begins entering the cells.
While on the Heat Station, your DNA is entering many cells. Now you need to trap the DNA inside. You do this by cooling the cells. At cooler temperatures, the cells’ membranes get more rigid which helps to trap the DNA inside the cells.
After 90 seconds have passed, you can move the tube back to the cold station where it should incubate for two minutes!
To help get your cells growing again, you’ll use Recovery Media. This is LB, a yellow liquid that contains sugars, amino acids, and minerals that E. coli love. Imagine your favorite food; Recovery Media is the equivalent for E. coli bacteria
The DNA Playground Hot Station is also used to incubate and grow your E. coli cells.
Your grown K12 E. coli cells are no longer Blank Cells! By putting the cells in Transformation Buffer and Heat Shocking them, you have caused a DNA program to enter into them, thus changing their genetic make-up. It’s time to get them growing again.
The Recovery Step is where you get your cells growing again. You add some yummy LB liquid, mix your cells, and incubate them.
Your Cold Station and your 42 ℃ Hot Station are both on. For the Recovery Step, you want to grow your cells at a comfortable 37 ℃. 37 ℃ is the ideal temperature to grow E. coli bacteria. Change the Hot Station from 42 ℃ to 37 ℃.
Grab your tube of Recovery Media and pour the liquid into your tube of cells and DNA. LB media is similar to the LB agar you used to make the LB agar plates. The difference is that there is no agar!
Impressive Pour! You’ve poured the Recovery Media into the DNA/Cells, but you need to mix them. Mixing is extremely important when bioengineering.
The best method here is to invert the tube 10 times, but you can also shake them.
You transformed Blank cells with a DNA plasmid that tells the cells to produce a colour pigment from coral. You added Recovery Media to help them start growing again. Now they need to grow at 37 ℃ for between 30 mins to 24 hours before you pour them on your selective LB agar plate that you made in the first step.
Drag your tube onto the 37 °C Hot Station.
Your transformed and recovered E. coli cells are now ready to be “plated” on the selective LB agar plate you made in the first step.
You’ll now use the selective LB agar plate that you made in the first step
You’ll use a sterile yellow loop to spread out the cells.
Pour your transformed cells onto the selective LB agar plate you made in the first step. Generally you only need to pour about 100 microlitres onto the plate, so pour half of your recovered cells.
Use the Yellow Loop to spread the Transformed Cells over the entire Selective LB Agar plate. You spread over the entire plate because unlike streaking blank cells where you start with millions of cells that can grow, after a transformation you only have tens to hundreds of transformed cells. In other words, streaking is not used in this step.
Next Step: Incubate Cells
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You’re done spreading the cells! You must now move onto the last step, Incubating Cells.
Just like when growing your Blank Cells, the Incubator will be used to grow your Transformed Cells
Your selective LB agar plate with spread transformed E. coli is ready to be incubated.
You’re ready for the final incubation step where you grow your engineered cells from individual bacteria to colonies.
Similar to when you grew your Blank Cells on non-selective LB agar, you need flip the selective LB agar plate with Transformed Cells.
Great! Over the next 24-48 hours your transformed bacteria will grow. Transformed cells grow slower than blank cells because they are now producing antibiotic resistance proteins and pigment.
Sometimes it can take 72 hours in order for the colours to really POP!
Watch Cells Grow
24 hour timelapse
You’ve genetically engineered E. coli bacteria to produce a Red Pigment from Coral!
7. Incubate Cells
Do the real, hands-on version of this experiment!
See what else is created using genetic engineering
You are now a Virtual Genetic Engineering Hero!
Do the real, hands-on version of this experiment!
Compare the transformed cells with the blank cells!
Diabetes is a medical condition that is often treated with insulin, but where does that insulin come from? Until the 1970s, insulin was collected from the pancreas of dead pigs and horses. The organs were dried then ground up, and the insulin (naturally produced in the pancreas) was isolated and injected into patients. This process helped many people control their diabetes but also led to allergic reactions and impure samples of insulin.
But in the 1970s scientists identified and isolated the DNA snippet that encoded human insulin! They put this snippet into a plasmid, transformed that plasmid into a bacterial cell and then cultured it – just as you did in this simulation. The engineered microbe was producing a safe human insulin: Humulin. In 2006, an estimated 150,000,00 vials of insulin were produced for millions of patients (ref).
Many of today’s medicines are produced with these bioengineering techniques. Bacteria and yeast cells are grown in large bioreactors to produce improved insulin, several cancer therapies, and other medicines.
Health & Medicine
Continue your journey
Learn about bio-art:
Canvas Kit simulator
Learn about DNA:
What is DNA? simulator
Test your genetic engineering skills with a quiz!
Do the real, hands-on
version of this experiment!
Learn something new:
Test your genetic engineering skills with a quiz
Learn about DNA:
What is DNA? simulator
Learn about bio-art: Canvas Kit simulator
Today, most pigments and dyes are made using industrial chemistry. This means lots of heat, energy, and in many cases, a significant impact on the environment and the health of people creating them.
Bioengineering is helping individuals and industries make inks, dyes, and pigments sustainably, from sources that were never before possible.
What if we could make a safer, better RED pigment?! It is your job to genetically engineer E. coli bacteria to produce a red pigment that comes from coral.
In this exercise, you will get a tube of DNA from your friend. This DNA is a circular string of DNA she developed, called a plasmid. When you insert it into bacteria, the bacteria will “read” the DNA and produce the colorful RED coral protein as the bacteria grows, a sustainable pigment!
Engineer Red Pigment
We need more sustainable RED !
Plants can be bioengineered too. Several food crops have been engineered to produce more nutrient rich, drought resistant, or pest resistant varieties. To engineer plants, a DNA plasmid is modified to express a specific function, such as vitamin production. The DNA is transformed into a bacterial cell, just as you did in this simulation. The plant is then mixed with this transformed bacteria and under special conditions, the DNA from the bacterial cell will enter the plant cells. The plant can then make the vitamin or whatever the DNA program directs.
The first transgenic plants were developed in the 1980s. These were antibiotic resistant tobacco plants. Today millions of people rely on genetically modified foods every day, but the technology remains controversial and people are concerned with the safety of foods in terms of human health and our planet’s ecosystem.
Until recently, the only way to make plastics was to produce them from fossil fuels. But these days, biology can be used to make materials like some bioplastics and biorubbers. Bacteria and yeast cells have been engineered with DNA that can build small carbon molecules inside the cells. These carbon molecules are the building blocks that can be connected together to make bioplastics and biorubbers.
When you think of energy, you probably think of coal or nuclear power plants. However, in recent years living organisms have been used to generate energy, usually in the form of liquid fuels.
Bacteria, yeast, and algae are engineered with new DNA to produce energy rich compounds: Attempts to produce a gasoline like molecule, butanol, in bacteria are underway. Ethanol can be produced in yeast cells for use in vehicles, and algae are engineered to produce kerosene, a component of jet fuel.
Researchers are also engineering bacteria to directly produce electricity - Look up microbial fuel cells to learn more!
Get New Loop
You touched the end of the loop!!! Now there are likely microorganisms on it. Get a new loop and please be careful next time.
Grow Blank Cells
A plasmid is a small circular piece of DNA. A plasmid typically has one or more genes that are the instructions that transform the cell. In this case, your plasmid has a gene for antibiotic resistance (selection), and for the creation of the red pigment.
Learn More About DNA
Objective: Become a genetic engineering hero!
Learn the basics of genetic engineering and how to engineering bacteria
1. Click on objects throughout, there may be hidden information
2. Ask any technical questions email@example.com or find
3. Don’t forget to download and print your Virtual Bioengineer Certificates!
You’re going to learn about the food and “houses” that scientists grow bacteria in (LB agar plates). You’re going to learn how to streak and grow cells (Grow blank cells). You’ll use chemistry to make your cells able to take DNA in (Make competent cells) and become engineered (Transform cells).
After you’ve engineered your cells, you need to recover them and grow them. In the end, you’ll hopefully get “colonies” of genetically engineered bacteria.
Have a look on the left side of the screen. You can see the seven steps that you will go through to engineer your bacteria to be “microfactories” that produce red pigment.
How does this work?
The hot station allows you to incubate tubes at warm temperatures. In labs, you would often use a hot water bath, and a thermometer.
This is your control panel for controlling your DNA Playground! You’ll press the buttons to control the machine!
The Incubator is a “little oven” in the side of your DNA Playground that can store two petri dishes (DNA Playground Home) or eight petri dishes (DNA Playground Large). The incubator can hold the right temperature for incubating E. coli cells and yeast cells
The cold station keeps tubes cold. In labs, you would often use ice in an ice bucket.
OK, got it!
Start the Engineer-it Kit Simulation!
No! I want to do it in real life
Day 1 bag:
LB agar powder, antibiotic, blank cells
Yellow & Blue loops
The Engineer-it Kit comes with all of the ingredients you need to do your first genetic engineering activity. This includes growing and engineering cells, and to safely inactivate (dispose of) your experimental waste. The kit contains the following items and you will learn about each of these during this simulation:
Day 2 bag:
T. buffer, recovery media, DNA, (+) cells
OK, Let’s Start
Sterile water (50 mL)
Petri dishes (4x)
LB Agar Powder
Start Making LB Agar Plates
As you do the simulator, click on objects. There might be hidden information!
Great Find! Good clicking!
Did you know that, despite popular belief, only a three types of E. coli bacteria out of hundreds of different types of E. coli are pathogenic? In fact, you have E. coli in your large intestine and they are helping to digest your food and maintain intestinal health.
K12 E. coli is a lab strain of E. coli that is not dangerious and has been used by scientists for more than 100 years!
This is your control panel for controlling your DNA Playground! Press the buttons to control the machine!
See the DNA Playground
Let’s start with a quick intro to the tools you’ll use in this Virtual Bioengineer simulation.
You’re going to use an “Engineer-it Kit” which has all the materials you need (cells, DNA, etc.) and a DNA Playground, which is the lab equipment for incubating and engineering your cells.
The Incubator is a “little oven” in the side of your DNA Playground that can store two petri dishes (DNA Playground Home) or eight petri dishes (DNA Playground Large). The incubator can hold the right temperature for incubating E. coli cells and Yeast cells
Its important to use sterile water when you bioengineer, because if there are any other microorganisms in the water, they can grow too!
LB is food for bacteria. Agar makes the LB jelly-like so you grow bacteria on the surface of it. LB contains sugar (glucose), tryptone (amino acids), and yeast extracts.
Petri dishes are the containers that you grow bacteria in. As you’ll see, you pour different kinds of molten LB agar into the dishes.
Antibiotics are added to molten LB agar to make the media “selective”. You need selective plates when you grow engineered bacteria.
When bioengineering you should wear gloves (latex, nitrile). You do this to protect you from your experiments and your experiments from you!
You need a microwave to boil your sterile water. Having boiled sterile water will make sure your LB Agar powder will dissolve
Put on your purple Nitrile Gloves. You should wear gloves to protect your experiments from you and you from your experiments
First, you must always label your plates! Grab the marker to label your plates so that it is clear what they are and who the plates belong to. It is an important part of biosafety to label your plates.
Ok! Let’s pour plates!
You must always label your plates on the bottom. That way if the lid falls off, the part that contains the LB agar will always be labelled.
You’re now ready to make your LB agar petri dishes.
LB agar is both the food that bacteria eat and a structure they grow on. LB agar is similar to Jell-O. If it helps, pretend you are making bacteria Jell-O!
Drag the sterile water into the microwave so you can boil it.
LB agar powder dissolves at ~80 °C, so your water needs to boil. Turn on the Microwave.
Your water is boiling!
Microwave a bit longer
Your water is not yet boiling! You need to see a rolling boil or your LB agar will not dissolve!
LB agar powder looks like Jell-O powder, but is more nutricious. It contains sugars, amino acids, and minerals that bacteria need to grow. Pour the LB agar powder into the boiling water.
You now need to swirl the LB agar so that all the powder dissolves into what is called “Molten LB agar”. In the lab, you might have to microwave the solution a bit more so that the liquid is hot enough to dissolve the powder. Microwaving for 5 more seconds after mixing is great practice.
At the end of this step, the solution will be slightly yellow, but transparent (not cloudy)
Pour non-selective plate
You now have Molten LB Agar. In the Canvas Kit you added antibiotics in the next step, however, in the Engineer-it Kit, you must first pour a “non-selective” plate before you add antibiotics.
You now need to add the antibiotic pill to your molten agar to make sure only your engineered bacteria grow on some plates. This is called selection (making selective plates). Drag the pill into the molten LB agar and gently swirl for 1-3 minutes until the insides of the pill dissolve.
After the antibiotic pill dissolves, you pour the molten LB agar into the three remaining plates so that each plate is half full. Then, you place the lid back on 3/4 of the way and let the agar cool. The molten agar will solidify. These plates will be used to grow your bacteria.
Put the lid on
Next Step: Grow Blank Cells
You are done with the first step! You’ve made LB agar plates that you will use to genetically engineer cells. Now, you are going to grow your Blank cells
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For the non-selective plate, you want to pour into the “bottom” (not the lid) of the Petri dish and fill it half full. As you’ll see, this plate will be extremely important for growing Blank cells.
Non-selective plates do not have any antibiotics in them. Therefore they do not select for any specific trait. These plates simply have the growth media (food) that bacteria love and will enable them to grow.
Selective plates have antibiotics added into them. These are called Selective plates because they help you SELECT for your ENGINEERED bacteria. The DNA that you’ll engineer your bacteria with has a gene in it to make your bacteria colourful, and also has a gene it in to make the cell resistant to the specific antibiotic you added to the molten LB agar.
By making your cells resistant to a particular antibiotic, your cells are the only ones that can grow in the selective plates; the non-engineered bacteria die.
Don’t worry, the antibiotic that you make the bacteria resistant to are no longer used in medicine/hospitals. They typically include ampicillin, kanamycin, chloramphenicol, and a few others.
Streak Blank Cells
A sterile inoculating loop is used to touch some cells that you received in the form of a “Stab" and streak them onto the LB agar. Careful not to touch the loop end. You’ll contaminate it!
Laboratory bacteria, also called cells, can be kept alive in many ways. The bacterial cells you are working with here are provided as a “stab”. Stabs named this way because the cells were stabbed into some soft LB agar in a tube. Bacterial cells that are grown this way can live for several months. You will transfer some of the cells from the stab to amplify them on a Petri dish for engineering later.
Laboratory E. coli (Escherichia coli) are not the same as the E. coli you might associate with bad hamburgers and infection. Lab E. coli don’t make healthy people sick. In fact, humans have lots of non-harmful E. coli in their guts to aid in the digestion of food and the creation of vitamins and amino acids. For this experiment, you’re using a well studied, non-pathogenic (not harmful) strain called K12. Almost every research and biotechnology lab in the world has used a version of this strain at some point in their research.
You did a great job making your non selective LB agar plate. Now its time to use it! Make sure you grab the NON-selective plate when growing “Blank” cells.
Start by turning on your Incubator to 37 ℃ so that it is warm when you need it
It can take up to 30 minutes for it to reach the desired temperature. Note that you can put your plates in the incubator even if the temperature is not yet reached. It is time to streak Blank cells.
Streak your Blank Cells
K12 E. coli live in the intestines of animals and like body temperature! It will take an extra day to grow them at 30 ℃. Turn on the incubator to 37 ℃.
Start by turning on your Incubator to 37 ℃ so that it is warm when you need it! Do you know why E. coli bacteria are grown at 37 ℃?
Do you want to waste electricity!?? The Cold Station uses 40W electricity, so why turn it on when you don’t need it? Turn on the incubator to 37 C.
K12 like E. coli live in the intestines of animals and like body temperature! Temperature of 42 ℃ is equivalent to a fever in a human body which is designed to kill microbes and puts stress on their survival! Turn on the incubator to 37 ℃.
Your Engineer-it Kit comes with a “Stab” of Blank Cells. You need to get some cells onto your loop so that you can grow them on your LB agar plates to grow them. Dip your sterile yellow loop into one of the stab of “bacteria paint” E. coli cells.
Whoa down there, you only need to dip once! Really, you only need to touch the surface of the LB agar stab because there are lots of cells there. If you get some jelly-like LB agar on your loop too, it is OK.
With your non-selective LB agar petri dish open, streak the surface of the LB agar with the bacteria that are on the loop.
If you successfully dipped your loop in the stab of cells, it will look “wet” and shiny when you have a quick look at it. If it looks dry, dip it again in the cells!
The goal of plate streaking is to rub cells onto the surface of the LB agar so that you grow bacteria colonies. A colony is a single bacterium that grows and divides into a small mound which contains millions of bacteria. Streaking plates involves Three steps:
1) Streak (drag) a line near the edge
2) With a new sterile loop, drag through the first line and do a zig
3) With a new or sterile part of the loop, drag through the zig and do further zig zags
Use the loop and streak cells on the second line, from 2A-2D.
Line 1 has LOTS of cells. By dragging a clean loop through line one, you’re getting less cells on Line 2. Think of it as diluting the amount of cells.
After you’re done, put the loop in the discard bin
Use the last loop and streak cells on the third line, from 3A-3F.
This line should have even less bacteria in it, including some isolated bacterial colonies after the cells have grown overnight.
Isolated colonies arise after a single bacterium divides many times to make a mound, which is what you want for a future step: transformation.
Put your streaked Petri dish onto your incubator paddle. Your incubator paddle will help you to insert your petri dish into your DNA Playground!
Put Humidity Chamber Ovover
Slide the Paddle under
Your Petri dish is on your paddle and it is now covered by the humidity chamber. It is now ready to insert into the DNA Playground!
Insert Into DNA Playground
Open the Incubator!
Insert the Petri dish
Let your cells grow for 12-24hrs!
Close the Incubator
You’re about to do an experiment where you need to lower the temperature to keep the cells stable! Turn on the Cold Station to 4 °C.
Good job! While this station is cooling down, you can get a tube of transformation buffer and put it in the cooling station.
Get Transformation Buffer
Cooling to 16 °C isn’t quite enough to keep your cells stable while making them competent. Cool it down further to 4 °C. 16 °C is often used in genetic engineering when ligating DNA (gluing it together).
You don’t need the incubator yet! Turn on the Cold Station so that you can make your cells competent and not kill them in the process.
You don’t need the hot station right now. You need to cool your T. Buffer and Blank cells, not heat them up! Turn on the Cold Station to 4 °C.
You don’t need the hot station right now. You need to cool your T. Buffer and Blank cells, not heat them up! Turn on the Cold Station to 4 °C.
Don’t turn off the Cold Station! Your cells will warm up beofre they are supposed to and your experiment will not work very well. Keep them cold until you’re ready to heat shock them.
You need to change the Hot Station temperature to 42 °C, not turn on the incubator! Turn on the Hot Station. You will use the incubator later!
You need to change the Hot Station temperature to 42 °C, not 30 °C!
Turn it up!
You need to change the Hot Station temperature to 42 °C, not 37 °C! Turn it up! You will use the 37 °C Hot Station during the recovery step.
Start the Transformation
Your DNA Playground stations are now reaching the right temperatures. You can now add your DNA to the competent cells while the Hot Station warms up.
Now that many DNA plasmids are bound to many cells, you need to “Heat Shock” the cells. By heating the cells from cold to hot the membrane of the cells becomes more fluid and the DNA is better able to pass through the cell membrane. This is a procedure called a Heat Shock.
Drag the tube of mixed cells and DNA from the Cold Station to the 42 ℃ Hot Station.
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Next Step: Recover Cells
After two minutes the cells are cold again, and the membranes are rigid. You’ve trapped many copies of your DNA plasmid inside many cells. The cells will now begin “running” the DNA programs in the plasmid!
You must now recover the cells!
You need to change the Hot Station temperature to 37 °C, not 30 °C!
Turn it down!
Great! Your Hot Station will cool to 37 °C. However, if you finish the recovery quickly and the Hot Station is above 37 °C you can still place your tube here because it will cool down shortly and will not harm your cells.
You do not need to press the Shock 42 °C button. Rather you can directly press the Heat 37 °C button to turn on the 37 °C Hot Station.
Great! Now incubate these cells overnight, up to 24 hours. During this time all the cells will grow and divide, including the cells that you engineered with the DNA plasmid.
In the next step you will plate your cells on the selective LB agar plates which will help you select for only the engineered cells.
You do not need the cold station anymore. You can turn it off.
Turn off Cold Station
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Next Step: Plate your Cells
Excellent, over the next hour, your incubator will slowly heat up to the optimal temperature for incubating your engineered cells.
Ooops, you turned on the 42 °C incubator! This is a bit warm for recovering cells. Instead, turn the incubator on to 37 °C.
Your cells would recover at 30 °C, however, not as well as at 37 °C. Because genetic engineering can be tricky, you want to have the best chance of success. 37 °C incubation is your best choice.
Let your cells grow for 24-72hrs!
Oops! These are the controls for the Hot station of the DNA Playground. The Hot station is to heat up tubes and is located on the top of the minilab. You want to turn on the incubator to 37 ℃. Incubators are made to incubate Petri dishes
You need to change the Hot Station temperature to 37 °C, not the make the cold station warmer!
You need to change the Hot Station temperature to 37 °C, not turn on the incubator!
You do not need to press the ice 4 °C button, it must stay on to keep your cells cold. You have to on the 37 °C Hot Station.