Topic 24: DNA Tools and Biotechnology

Question 1

Compare and contrast Sanger sequencing and next-generation sequencing. Draw a ddNTP. How are ddNTPs used in DNA sequencing?

Answer 1

Sanger sequencing sequences a single DNA sequence at a a time and Next-generation sequences millions of fragments at a time(there are so many reads)
Sanger use a primer: is specific
Next generation uses a universal adapter: not specific

Question 2

Explain the process of Molecular Cloning? What are restriction enzymes? What kind of ends do they produce? Why does this matter?

Answer 2

Molecular cloning a way to create identical copies of a DNA segment.
Restriction enzymes cut DNA molecules at a specific DNA sequence called restriction sites.
They can produce blunt ends or sticky ends
Can allow another DNA segment to enter and bind in.
DNA ligase glues them back together

Question 3

Describes the steps in PCR and gel electrophoresis.

Answer 3

Steps in PCR:
- produces many copies of a specific fragment of DNA.
- USES A THREE STEP SYCLE:
- Heating: Heat used to Denature
- Cooling:
-Replication:
Usually a 3-hour process

Gel electrophoresis: when we’re looking for DNA.

Question 4

Describe the tools that scientists would use to study the expression of a few genes

Answer 4

Tools scientists use to study the expression of a few genes
- in situ hybridization: allows us to look at mRNA in real life.
- RT-PCR: reverser transcription: conveys mRNA to DNA, uses PCR to amplify the DNA, and it runs on a gel.

Question 5

Describe the tools that scientists would use study global gene expression (transcriptome analysis)

Answer 5

Tools used to study global gene expression:
By using microarrays- they contain thousands of thousands of dot of DNA/ Each dot contain DNA sequences.

Question 6

Describe the tools that scientists use to study gene function.

Answer 6

Crispr-cas9 used to investigate gene function by direction editing the gene of interest. GWAS to learn which SNPs are associated with disease.