Day

Approximate time used

Content

1

1 h

Introductory remarks on:

biosafety in the handling of chemical and biological materials

procedures on DNA extraction protocols

2:30 h

DNA extraction solutions preparation and autoclaving

30 min

Blood sample collection

1:30 h

Protocol 1 starting. Hemolysis and nucleus lysis, overnight

1:30 h

Protocol 2 starting. Hemolysis and nucleus lysis, overnight

2

3:30 h

DNA extraction using Protocol 3

2 h

Protein extraction and DNA precipitation of samples from protocols 1 and 2

3

2 h

DNA recover, wash and solubilization of samples from protocols 1 and 2

1:30 h

Spectrophotometric quantification of DNA extracted from protocols 1, 2 and 3

2:30 h

Agarose gel electrophoresis of Genomic DNA

4

4 h

PCR reaction

2 h

Agarose gel electrophoresis of PCR products

5

4 h

Graphics and Statistical analysis of the results to answer the questions above:

I. Using spectrophotometer quantification calculate mean and standard deviation of DNA solutions:

a) Concentrations

b) 260/280 ratio

c) Yield per protocol

d) Yield per blood volume used

II. Construct graphics using the calculated values for a, b, c and d items

III. Identify the statistical difference among the three DNA extraction techniques on a, b, c and d items