Omics, Tools and Techniques

Question 1

seperating DNA

Answer 1

1. blend sample with salt water
2. emulsify with a detergent
3. centrifuge to seperate large solids.
4. add alcohol to precipitate dna as a solid.

Question 2

Sanger sequencing or (synthesis sequencing)

Answer 2

a DNA synthesis reaction using terminator nucleotides - dideoxy-NTPs

• wherever a ddNTP is incorporated the synthesis stops.
• Large numbers of variable sized DNA fragments
• If you make enough, you end up creating DNA fragments that differ in length by one nucleotide.

Question 3

PCR - Polymerase Chain Reaction

Answer 3

The process of turning a single DNA molecule into billions of copies using a few simple reagents and a heat source.

Question 4

PCR steps (5)

Answer 4

To the DNA of interest, add:

1. synthetic short ssDNA “primers” - oligonucleotides
2. thermostable DNA polymerase - Taq polymerase
3. deoxy-nucleotide triphosphates - dNTP
4. heat to denature the DNA 95C
5. Allow complimentary base pairing to bind primers - 45-60C
6. Allow DNA synthesis with Taq Polymerase