NAME: Ang, Antivola, Silva, Te, Vizcarra GROUP: 2 –Targaryen

TITLE OF ACTIVITY: Central Dogma of Molecular Biology DATE: 19 September 2017

Objectives
1. To devise a three-dimensional model of DNA
2. To examine DNA replication and protein synthesis through RNA transcription and translation
3. To find out the correct nucleotide base pairings and associate these with genetic mutation
4. To draw out DNA from own cheek linings and various fruits
5. To visually distinguish actual observations of DNA fragments from DNA yield

Materials to be Used
Part 1: 3D DNA Model for Replication, Transcription, and Translation
- Clay (different colors)
- Bamboo sticks

Part 2: Extraction of DNA from a Fruit Part 3: Extraction of DNA from Buccal Cells
- Banana, kiwi, and strawberry - 5 mL of 25% liquid detergent
- 3 resealable plastic bags - 25 mL clear centrifuge tubes (1 per student)
- Plastic knife - 10 mL of 0.9% salt water
- ½ spoon of salt - Cup (1 per student)
- 20 mL of warm water - 1 teaspoon of 5 mL chilled alcohol
- 3 plastic cups - Barbecue stick (1 per student)
- Clean strainer/paper towel/cloth - 2.5 mL microcentrifuge tubes with cold
- 1 spoon of liquid soap alcohol (1 per student)
- 20 mL of cold alcohol
- 3 sticks
- 3 clear centrifuge tubes (20 mL)
Procedures
Part 1: 3D DNA Model for Replication, Transcription, and Translation

Clay, bamboo sticks – create 3D DNA model with 9 nucleotide base pairs

Use corresponding colors, shapes – properly label molecules

Model is standing, twisted double helix

Adjust model – apply correct nucleotide base pairing (complementary strands)

Make model of single strand DNA to mRNA sequence: AUGGUGUUAUAG

Find tRNA and corresponding amino acid using table

Part 2: Extraction of DNA from a Fruit

Peel banana

Cut

½ cup in resealable bag

+ ½ spoon salt, + 20 mL warm water

Squash

Filter to plastic cup with strainer

+ 1 spoon liquid soap

Stir gently

+ 20 mL cold alcohol

Wait 5 minutes
 Collect fragments with stick

Transfer to vial with 5 mL cold alcohol

Label vial

Observe
Part 3: Extraction of DNA from Buccal Cells

5 mL of 25% liquid detergent to centrifuge tube

Gargle 0.9% salt water for 30 seconds

Spit into cup

Pour 10 mL buccal cell mixture to centrifuge tube

Close and mix 2-3 minutes without bubble formation

Open and pour 1 teaspoon chilled alcohol

Set for 5 minutes

Collect DNA fragments by swirling barbecue stick

Place in 2.5 mL microcentrifuge tubes with cold alcohol

Data
Part 1: 3D DNA Model for Replication, Transcription and Translation
Original DNA
DNA Sequence:
Strand 1: ATG GTG TTA TAG
Strand 2: TAC CAC AAT ATC
DNA template
Sequence of DNA template:
ATG GTG TTA TAG
Initiation
What is tRNA sequence and
amino acid of the start codon?
1st: UAC - Methionine
REPLICATION
Separation of DNA strands Complementary strands
What is the enzyme responsible Complementary strand
for separation of the strands? sequence:
Helicase Compl. strand 1: ATG GTG
TTA TAG
Compl. strand 2: TAC CAC
AAT ATC
TRANSCRIPTION
mRNA transcript mRNA Codons
What are the four codons
AUG GUG UUA UAG from the mRNA transcript?
AUG – GUG – UUA – UAG
TRANSLATION
Elongation Termination
What is tRNA sequence and What is tRNA sequence and
amino acid of the second and amino acid of the fourth codon?
third? 4th: AUC – Stop
2nd: CAC – Valine
3rd: AAU – Leucine Translation stopped. What do
you call this codon? Stop Codon
Parts 2 and 3: Extraction of DNA from Fruits and Buccal Cells
Insert picture of the corresponding steps from your experiment. Write observations.
Strawberry Banana Kiwi
Mashing of fruits

x x

Observations: Observations: Observations:

x - ½ spoon salt, 20 mL water x
added before mashing
- Some bubble formation
manifested
- Mashed banana became
watery
Addition of detergent

x x

Observations: Observations: Observations:

x - Mashed banana was strained x
before putting liquid soap
- Mixture became greenish in
color after gently stirring
- Some bubble formation
manifested (due to soap)
Addition of cold alcohol

x x

Observations: Observations: Observations:

x - Layer formation noticeable x
even without mixing yet
- 5 minutes setting time done:
layers were distinctly
apparent
- Clustered particles (DNA of
banana) float on alcohol
layer
Final output

insert picture

Observations: Observations: Observations:

- Yielded DNA from - Amount of DNA yielded - Yielded DNA from kiwi is
strawberry is most apparent from banana is large minute
among banana and kiwi - DNA clumped together - DNA did not cluster to form
- Large clumps acquired clumps
 Collected buccal cells Addition of buffer
Observations: Observations:
- Bubble formation is apparent as member spits - Some bubbles stayed on top as 25%
the gargled 0.9% salt water dishwashing liquid was added
- No DNA yielded yet - Dots and strands started to form on top

ANG ANTIVOLA SILVA TE VIZCARRA

Final output
Observations:
- White dots and strands (DNA fragments) became more apparent on top
- Using a stick, fragments were transferred to a vial with alcohol
- Each member yielded distinct amounts of DNA
- Ang: member with the most strands or fragments to appear
- Antivola: strands clumped into a single “ball”
- Silva, Te, Vizcarra: some strands apparent