Cell dynamics: extraction of Dna, qualitation + quantification of dna

What is a supernatant

Contains the lysed rbcs

What is an enzyme that degrades Dna

Dnase

Function of TE buffer

To prevent Dna degredation, used to store Dna

Function of Edta in TE buffer

The Edta is a chelating agent. It chelates Ca and Mg ions which are cofactors required by Dnases

Where is Dna found in blood

Leucocytes (wbc)

How was dna extracted from blood leucocytes

The leucocytes are isolated using a hypertonic solution(red blood cell lysis solution)

The lysis of the leucocytes occurs using sds to gently disrupt the cell membrane(white cell lysis solution)

When placed in an electric field where does Dna migrate

From the negative cathode to the positive anode

What is gel electrophorisis

Gel system (e.g agarose) used to separate Dna according to size

Is the phosphate and sugar backbone in Dna soluble of not soluble in water

Soluble

The electrophoresis gel is stained with gel red, what was used prior to this

Ethidium bromide

In the process of isolating Dna, how were proteins and contaminants removed

Using a high salt solution for ‘precipitation’ by ‘salting out’ proteins (10M ammonium acetate was used)

In the process of isolating Dna, how was dna recovered and washed

Isopropanol used to precipitate out insoluble Dna, washed using 70% ethanol

Function of nanodrop

Used to measure concentration no dilution needed

Calculation for dna concentration

A260(50)(dilution factor)/1000=conc in micrograms/microliter