Henrietta Lacks: Page 2/10

Molecular biology techniques are being increasingly used in diagnostic laboratories.
The polymerase chain reaction (PCR) is one of the newest techniques available and potentially offers tremendous advantages for the diagnosis of infectious diseases.

Human papillomavirus infections cannot be easily diagnosed by the conventional methods used in the virus diagnostic laboratory, but PCR can be used for the detection of these viruses.

In the laboratory, perform the following experiment:

You are provided with the following reagents:

Sterile distilled water
HeLa cell DNA
MRC-5 cell DNA (a human diploid cell line)
Sequence-specific synthetic DNA oligonucleotide primers for human papillomavirus type 18 (HPV18)
10X PCR Buffer
Taq DNA polymerase
Mineral oil

Label 3 small Eppendorf tubes as follows:

N (negative control)

H (HeLa)

M (MRC-5)

Carefully, without cross-contaminating the reagents, to each tube in this order add:

9µl Sterile distilled water

2µl 10X PCR Buffer

2µl Oligonucleotide primer pair

5µl of the appropriate DNA (water for negative control)

2µl Taq polymerase

Overlay each reaction mixture with 50µl of mineral oil.

	N (negative control) H (HeLa) M (MRC-5)
Carefully, without cross-contaminating the reagents, to each tube in this order add:
9µl Sterile distilled water 2µl 10X PCR Buffer 2µl Oligonucleotide primer pair 5µl of the appropriate DNA (water for negative control) 2µl Taq polymerase
	Overlay each reaction mixture with 50µl of mineral oil.

In the laboratory, perform the following experiment:

Put the tubes in the automated thermal cycler ('PCR Machine')