Tools/Sequencing

Question 1

Why is CRISPR so Bad @$$?

Answer 1

It has RNA guided endonuclease activity (cas9).

This allows for genomic engineering and

Question 2

What are the 3 components of a cloning vector?

Answer 2

1. Restriction cut site
2. origin of replication
3. Selectable marker (amphicilin resistance gene)

Question 3

What are micro array analysis used for?

Answer 3

Analyze expression of thousands of genes at the same time.

Question 4

How does Sanger sequencing work?

Answer 4

Add one DI-deoxy nucleotide at a time to halt replication. Then you compare strands by electrophoresis

Question 5

What disease can be identified using RFLP?

Answer 5

Sickle Cell Anemia. It has a mutation that lies in a restriction site.
Steps to analyzing
1. Use Restriction enzyme to cut
2. Electrophorese
3. Southern Blot

Question 6

How does DNA fingerprinting work?

Answer 6

1. PCR with primers that surround variable number tandem repeat
2. Electrophoresis
3. Compare sample against target(s) for similarity

Question 7

Describe the basic process of PCR

Answer 7

Add ingredients (Taq Pol, nucleotides, primers)

1. Heat (denature DNA)
2. Cool (Anneal primers)
3. War, (activate TaqPol)

Question 8

What is the advantage of long read sequencing?

Answer 8

They can link variants together in a haplotype–providing contiguity of variation in blocks.

Question 9

What is Exome sequencing and how is it used?

Answer 9

Mapping the “Exon” part of the genome. Used to identify single nucleotide polymorphisms in patients with same genetic disorders.

Question 10

What are the characteristics of a disease that are likely diagnosable with exome sequencing?

Answer 10

1. Single Gene mendelian
2. Variants that are RARE and not in any database
3. Variants are non synonymous (change in amino acid residue). These have functional consequences

An example is Millers Syndrome, a disease caused by embryonic exposure to methotrexate.