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:
Label 3 small Eppendorf tubes as follows:
- 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
N (negative control)
|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.
© AJC 2007.