INTRODUCTION

(Title slide on screen) (Beaker drawn on the whiteboard > off to the right side)
(DNA model (centre) and cell models (right) on the floor, BIG X behind desk)
Under table: A box with beaker of crushed strawberries, test tube with diluted DNA sol.
(Stand in front of desk) Left to right (facing the audience) : Kaze, Celine, Abby, Katie
Kaze: I wonder why we’re all different…

Katie: This is because of…

Celine: DNA!!!

Abby: I’m Abby, this is Kaze, Celine & Katie. We’re team 16 from Singapore International
School and today we will be unveiling the mystical world of DNA.

Kaze: Our theme is to reveal the invisible, and so today we shall show you what actual DNA
looks like!

Katie: Hold on, aren’t DNA so small that it cannot be seen using the naked eye? Also, what
is DNA?

Celine: To answer these questions, we will be performing a simple experiment.

Abby: But first let us see what DNA actually is!

WHAT IS DNA
(Celine and Abby lift DNA model)

Kaze: This is the DNA. It is made of poly nucleotides that are joined together by hydrogen
bonding. ​You can see that all the pairs are made up of the same colour pairings. This is
because complementary base pairings create the hydrogen bond mentioned. The adenine
and thymine will always pair up whilst the cytosine and the guanine always pair up.

(Kaze moves to computer to prep slides)

Katie: ​Along here, this is what we call the backbone of the DNA. Although not shown, it is
made up of alternating sugar and phosphates.

Celine: It is also in a double helix structure due to the allowed bond angles in the
sugar-phosphate backbone, the way the bases stack together and the interaction between
the two strands.

Abby: Here is an animation that illustrates the structure of the DNA.

(Kaze clicks through slides)​ ​(Nucleotide animation on screen)
The bases are hydrophilic - meaning that they’re soluble in water - due to the negatively
charged phosphate groups. As you can see, the asymmetrical nucleotides stack up on each
other in a staggered manner, but with just one strand, the bases are still in contact with the
water in the cytoplasm. To prevent these bases from dissolving in the water, another strand
come and the bases of each bond with their corresponding bases. Finally, to prevent the
atoms from bumping into each other, the 2 strands twist to form a helical structure.
(place DNA model on the floor)

Celine: So what is the use of DNA? And how are they related to jeans? ​(fling jeans out)

Abby: Not jeans… genes! Genes are units of the genetic information. They regulate and
code for how the amino acids are arranged to form polypeptides, and can also regulate how
other genes are expressed! The order of the nucleotides make up the genetic code.

Kaze: But how are all of our cells different? Do we have different genetic information in all of
the cells? And how is this related to DNA? ​(Walk back to position behind desk, switch screen
to ​camera: pointing to big beaker​)

Katie: Well, all cells of an organism contain the same genetic information however different
parts, in other words different genes, are expressed in different cells to allow for cell
differentiation. You can think of the gene as a piece of information whereas the DNA is the
material that’s storing the information.

Celine: That’s so cool that something so tiny contains all the information of a living organism!
Is there a way to see them though? How would we be able to get them out of the cell?
(Walk back to behind the desk)

Abby: Well, let’s start the experiment to see DNA! ​(point dramatically at desk)

EXTRACTION
Kaze: Now before we start anything, we will need to prepare 200ml of 99% ice cold isopropyl
alcohol. Due to time constraints, we have already placed the 200ml of isopropyl alcohol in
this ice box. ​(gesture at ice box on the right end of table behind abby & kt)

(Kaze hold up strawberries while placing them in mortar)

Katie: We will be extracting the DNA from strawberries because they’re soft enough to be
crushed without a blender, and c'mon, who doesn’t love strawberries?

Celine: Here, to speed things up, we’re gonna pass the strawberries down so please help
with the process of crushing it.​ (walk to judges)

Abby: You can use any way to squish it so long as it all turns liquidy and jelly-like. This
increases the surface area of the strawberries to maximise the exposition to the extraction
solution later.
(Abby and Katie prepare models)

Kaze: On top of that, this step physically destroys the cell wall.
(Gesture at Abby who will point at Cell model)
Here, you can see that the cell wall is the outermost layer and the first obstacle we
encounter to get to the DNA.

(Abby puts down cell model, helps Katie take contents out of box)
To magnify what is happening in the cells, we have prepared a model. To demonstrate the
removal of the cell wall, we shall remove the cell wall from our own model.

Katie: Now, it’s time to prepare the most important thing: the extraction solution.
(point dramatically at desk)
(Abby and Katie prep whiteboard and “ingredient” labels)

Celine: ​(Celine talks while pouring)

To make it, we’re going to pour 180 ml of water ​(“water” on board) ​and 20 ml of unscented
soap ​(“soap” on board)​ into this huge beaker. Wait, Abby, what’s the soap for?

Abby: Well, first, we need to know its structure well and remember that cell and nuclear
membranes are made of phospholipids.

(“soap and phospholipid” on board)

(Katie prep model, Abby prep balloon and sharp object)
Kaze: As you can see, soap is actually similar to the phospholipid bilayer structures,
although not identical, the soap will interrupt the phospholipid bilayer structure and damage
the cell and nuclear membrane.

Katie: ​(Points at Cell Model) ​See here, the cell and nuclear membranes are the next layers
to get through and this is done by the soap like this. ​(Point at Abby who will pop balloon)
After these barriers to the DNA break, now, theoretically, the DNA can be released from the
cell.

Celine: We are now going to add 1 teaspoon of protease ​(“protease” on board) ​into the
beaker. ​(Celine talks while adding)

Abby: Though absent from the earlier DNA model​, there are actually many proteins
associated with DNA, such as histones.

Katie: Here, let’s magnify it so you can see! ​(Walk out from behind desk with BIG X)

Kaze: Histones are a type of protein, so the protease . They are wrapped around twice by
the DNA strands due to this attraction.

Celine: By adding the non-iodized salt, the sodium ions will displace a lot of these proteins
like this.

Abby: Last but not least, we add a bit of salt into a beaker.
Kaze: The negatively charged sodium ions in the salt will neutralize the positive charge of
the DNA's sugar phosphate backbone when the isopropyl alcohol is added. This allows the
DNA to be precipitated later.

Katie: Judges, I hope the strawberry you were crushing is ready.

Abby: ​(talking while collecting) ​Because now that you know what will happen with the
solution, it’s time to see for yourself. …. Thank you!

Celine: ​(Take big beaker of crushed strawberries from box)

Due to the time constraint, luckily we prepared some strawberries beforehand. Don’t worry
judges, your hard work will not go to waste. ​(Celine adds small amount into big beaker)

Kaze: After they’re all squished like this, it’s time to add the extraction solution into the
strawberry puree. ​(Kaze pours extraction solution into strawberry puree, mix)

Katie: While they do that, can anyone predict what we’re going to see? … ​(entertain
audience)​ HAHA that was a trick question!

Abby: You won’t see anything because everything that we explained before is happening
inside the strawberries.

(Kaze & Celine done mixing)

Celine: Yep, there’s a lot going on to remove all the barriers protecting the DNA. Okay, now
that all the membranes are damaged and the water-soluble DNA is freed, how do we extract
it out of the water?

Kaze: Hold your horses, before we extract the DNA, we have to filter the solution to remove
the cell debris. ​(Kaze and Celine filter solution)

(Katie walks to right end of table to take alcohol, pour into measuring cylinder 200ml?)
Katie: Meanwhile, we’ll take out the cold 99% isopropyl alcohol prepared earlier. Later, we’re
going to mix the strawberry liquid with this alcohol to cause the DNA to precipitate.

Abby:This needs to be cold because first, lower temperatures promote the flocculation of the
nucleic acids so they form a larger complex.

(Kaze & Celine done filtering)

Celine: It also prevents fragile hydrogen bonds between the nucleotides in the DNA from
breaking. Everything to make our resulting DNA similar to its original form.

Kaze: Also, so as to not keep you waiting, this large temperature difference lowers the
energy of the DNA faster, allowing them to precipitate quickly. Now, let’s get to it.
(Mix alcohol with solution → pause for a while)
Hey Katie, since it’s taking a while, why don’t we talk about why DNA precipitates in alcohol.
(Katie in front of desk again with Abby)
Katie: Sure! DNA is soluble in water, because like water molecules, they’re polar and can
electrostatically interact with each other thanks to their hydrophilic phosphate group.

Abby: ​(Thinker pose facing Katie) ​But didn’t we add salt earlier? I thought the positive
sodium ion will go with the phosphate group and neutralize it. Shouldn’t this make DNA less
soluble in water already?

Katie: ​(Finger guns @ Abby) ​Touche, but too bad water has a high dielectric constant, it
prevents this neutralization process. That’s why we’re using isopropyl alcohol with its low
dielectric constant!

Celine: Hey guys! Take a look, can you see the white stringy stuff floating to the top?
(Katie and Abby excitedly run back to look, may point camera for better angle
Abby: Ayeee, this is the DNA!

Kaze: So now the last step is to filter the DNA from the solution and wash it with isopropyl
alcohol.
(Celine holds the tea bag whilst Kaze pours DNA into tea bag to filter DNA, Katie sets up
spectrophotometer, Abby move camera to set up)
However, due to time constraints, we have prepared a sample of the DNA this morning.

(pass DNA to judge after done filtering, maybe talk while walking)
Katie: And there you have it! We have revealed the normally invisible DNA.

Katie: Of course, you should not believe everything we tell you. How do we know if this is
actually DNA? To prove it, we will be testing the purity of the DNA using the
spectrophotometer.

Abby: ​(Point to screen) ​Ta da! ​Here is the set up. For this we need DNA in aqueous state, so
to save time, we prepared it beforehand with a dilution factor of 100.
(Abby pulls DNA solution from box, Katie goes to write ​formula on board​ at the side)

(Celine & Abby filling cuvettes)

Kaze: Here is the formula we use to calculate its purity.
(Katie wheels in whiteboard and points)
You see, we need to know the absorbance of the solution at the wavelengths 260 nm where
DNA absorbs light most strongly, 280 nm and 320 nm.

Katie:​(Abby & Katie switch spots, K holds the two cuvettes up) In this cuvette, we have the
DNA solution. In the other, we poured pure distilled water. So first, the water is used to
measure 100% transparency. Then, to measure absorbance at other wavelengths… ​(Katie
and Kaze beep beep spectrophotometer)

Celine: At 260 nm, we have ____. And at 280 nm, the reading is _____. Finally, at 320 nm, it
is ______. ​(Abby write readings down at board)​.
Abby: So after correcting for turbidity ​(subtract A320 reading)​, we can take the ratio between
the reading at 260 nm and 280 nm and compare with literature value. ​(Spin board around to
calculate and write answer)

Kaze: If we get a value between 1.7 to 2, that’s a sign for good quality DNA. Time for the
moment of truth! …. ​(Abby spins board around) ​So, we got ____.

Katie: Well, there are many possible reasons why our result doesn’t match.

Over range

Celine: Firstly, we need to know that pure RNA should have 2.0 as its value. So this could be
a case of RNA contamination. This can be solved by using RNase to remove RNA during
purification.

Under range

Celine: Protein and some other contaminants may absorb strongly at 280 nm so this will
lower the value. Salt can also be one of them! Some may have been left in the extract since
isopropyl alcohol is also very insoluble to salt. This can be solved by using 70% ethanol to
wash off the salt after the DNA solidifies.

Abby: But it’s all good because at least we got to reveal the normally invisible DNA. As much
fun science is to do on our own, our equipment and procedure is not all that accurate when
compared to professional researchers.
(not too sure about the ending) 
 
Celine: Still, don’t you think it’s crazy that we can’t see some of the most important
processes in our body? Science *pop balloon* provides us a way to magnify and reveal
*unfold paper inside* all this and today we showed you one method.

Abby: We hope you continue on to find out about other things we don’t see or understand
after our presentation.
All: *flip paper: THANK YOU in huge letters* Thank you for listening!