DNA extraction lab report By Y10J Chan Yin Wing Stacey

Title(Bi) Effect of ethanol (Independent variable)on strawberries(Dependent variable)

Research(Bi)
The aim of the experiment is to investigate the effect of ethanol on strawberries.
Different masses of ethanol were added to the strawberries and DNA was extracted
afterwards for measurement.
Scientists extract DNA for a variety of reasons. First, it gives scientist a closer look of
the cell so as to identify it. By observing and studying DNA, scientist could also identify
genetic diseases and even create a vaccine against it. Moreover, DNA extraction makes
the classification of organisms easier. Furthermore, DNA extraction can enhance
genetic engineering in order to help people with diabetes.
In addition, equipment such as detergent, salt and alcohol was used during DNA
extraction.Detergent is used to break down the cell membrane and so DNA could be
released from the cell.Besides, salt is needed to remove proteins along with solidifying
and making DNA visible.Moreover, alcohol is also required in DNA extraction in order to
pull water from the DNA molecules so it will collapse on itself, leading to the
precipitation of DNA .
Hypothesis(Bii)
I predicted that the mass of the DNA will be affected by the mass of the ethanol
added.The reason I have this prediction is because according to my research, alcohol is
used to force the DNA to precipitate in the solution.Thus, I predicted the more ethanol
added to the solution, more DNA can be extracted.

Variables(Biii)
Independent variable The mass of ethanol added

Unit ml

Explanation Different masses of ethanol were added to the the

strawberries. 5ml,10ml,15ml and 20ml of ethanol were
added respectively into the strawberry solution.

Dependent variable The mass of DNA

Unit grams

Explanation It is measured by the trial balance.

Control variables Unit Explanation

water ml Water was controlled at 90ml.This is because it helps to keep

DNA intact during extraction.

salt ml Salt was controlled at 1.42g. As mentioned in my research, it

helps to solidify and make the DNA visible.

detergent ml Detergent was controlled at 10ml. It is mentioned in my

research that detergent breaks down the cell membrane so
DNA could be precipitated.

strawberries gram 3 g of crushed strawberries were used each trial and were
separated into 4 test tubes with different masses of ethanol
added afterwards. This is because mass of DNA depends on
the mass of the strawberries. Thus, the mass of the
strawberries used in each trial should be the same.

Equipment(Biv)

-1 zip-closer sandwich bag

-1 cheesecloth
-225 ml of ethanol
-4.26 g of salt
-270 ml of water
-30 ml of detergent
-1 teaspoon
-20 strawberries
-3 1x100 beaker
-4 test tubes
-1 test tube rack
-1 trial balance
-1 dropper
-1 knife
-1 pair of forceps

Risk Assessment(Biv)
Safety issues Safety precautions needed Emergency

Ethanol is highly We need to handle it with care We should use a fire

flammable. and avoid it from fire. blanket it cover the item.

Students may get We should cut the strawberries We should go to the

injured when using a slowly and carefully. medical room immediately.
sharp knife to cut the
strawberries into
pieces.

Detergent is irritant. We should handle it carefully We should wash our hands

and avoid it from touching our in running water if
skin. detergent is stained on it.

Method(Bv)

1. The strawberries were cut into small pieces and placed inside the sandwich bag.
2. The air from the bag was removed and the the strawberries were compressed.
3. The bag was crushed against the table and a strawberry slurry was formed.
4. The strawberry slurry was poured into a 100ml beaker.
5. 90 ml of distilled water,10 ml of detergent and 1.42 ml of salt were mixed in
another 100 ml beaker.
6. The mixture was added to the strawberry slurry and the solution was stirred.
7. A two-inch square of cheesecloth was placed over another 100 ml beaker.
8. The solution was poured over the cheesecloth,which separated the large solids.
9. The liquid and DNA that passed through the cheesecloth was collected in the
beaker.
10. By using a dropper, the liquid was transferred to 4 test tubes with equal masses
of strawberry liquid.
11. By using a dropper, 5 ml,10 ml, 15 ml,20 ml of cold ethanol were added to the
test tubes respectively.
12. The DNA was precipitated from the liquid.
13. By using a pair of forceps ,the DNA was extracted from the test tubes and was
placed onto the trial balance for measurement.
14. The mass of DNA in each test tube were measured in gram and recorded.
15. The experiment was replicated twice.

Results(Ci)

The mass of DNA extracted by adding different masses of ethanol

Mass of ethanol
added(ml)
Trial 1 Trial 2 Trial 3 Average

5 ml 0.13 g 0.12 g 0.09 g 0.11 g

10 ml 0.14 g 0.15 g 0.11 g 0.13 g

15 ml 0.16 g 0.16 g 0.13 g 0.15 g

20 ml 0.19 g 0.17 g 0.16 g 0.17 g

25 ml 0.20 g 0.19 g 0.18 g 0.19 g

Graph (Ci)

Conclusion(Cii)
Referring to my graph, the mass of DNA extracted increased with the mass of ethanol
added.Besides, according to my results, the by adding 5 ml,10 ml and 15 ml of ethanol,
 an average of 0.11, 0.13 and 0.15 was extracted respectively. The reason why this
happened is because the use of ethanol forced the DNA to precipitate. DNA does not
dissolve in water. Thus, with the use of ethanol, water is pulled from the DNA molecule
so that it will collapse on itself. This forces the precipitation of the DNA and also makes
it visible. Hence, the more ethanol was added, the more DNA is precipitated out if the
solution. In other words, the more ethanol added to the strawberry solution, the more
DNA can be extracted.

Conclusion(Ciii)
Referring to my hypothesis,I predicted that the mass of the DNA will be affected by the
mass of the ethanol added. According to my results, the mass of DNA extracted
increased with the mass of ethanol added.Therefore, my hypothesis was correct.

Evaluation(Civ)
The method of adding different masses of ethanol to the strawberry liquid is valid and
the results should also be reliable.However, the reliability may be affected by the
following issues.First, at the beginning of the experiment, the strawberry solution was
poured into a cheesecloth and was squeezed in order to separate the large solids. The
separation of strawberry liquid from large solids is valid to let the DNA pass through for
measurement. However, if the cheesecloth was squeezed too hardly, some of the DNA
may be damaged. As a result, the mass of the DNA extracted may be decreased.
Secondly, the difficulty in controlling the dropper may lead to uneven drops added to the
test tube. This will strongly affect the mass of the DNA obtained and the reliability of the
results. However, the experiment was replicated twice in order to increase its accuracy
and the reliability of the results. Fortunately, the results of each trial were similar and
were matched to my hypothesis. Therefore, my method is valid, accurate and reliable.

Evaluation(Cv)

Problems with the The effect caused by the Suggested

experiment problem improvement

When the strawberry solution Since the DNA is damaged,when The cheesecloth should
was poured into the ethanol is added, less DNA will be squeezed gently in
cheesecloth and was be precipitated from the solution. order to separate the
squeezed, some of the DNA This will affect the mass of the large solids so the DNA
may be damaged. DNA extracted. could pass through.

The strawberries used may be This will affect the mass of DNA The size of the
different in sizes and masses extracted when ethanol is added. strawberries used
so they might contain should be the same.
different masses of DNA. Besides, the masses of
the strawberries should
be measured at the
beginning of the
experiment.

The DNA may be too tiny
which makes it difficult to
extract it from the test tube for
measurement.
If all the DNA is not fully
extracted ,it may affect the
accuracy of the results.
I should wait for a few
minutes to let the DNA
solidify. In this way, a
thicker layer of DNA
could be extracted for
measurement.

Bibliography

"DNA Extraction." DNA Extraction. N.p., n.d. Web. 26 Sept. 2016.

"Work of Salt Isopropanol and Ethanol." N.p., n.d. Web. 27 Sept. 2016.
<http://info.gbiosciences.com/blog/bid/156468/Work-of-Salt-Isopropanol-and-Ethanol-in-DNA-
Extraction>.
@drnickoswald. "Ethanol Precipitation of DNA and RNA: How It Works - Bitesize Bio."
Bitesize Bio Ethanol Precipitation of DNA and RNA How It Works Comments. Bitesize Bio, 20
Sept. 2013. Web. 26 Sept. 2016.