User:Saroj Pandey/Notebook/SNP PCR optimization/2014/09/17

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gDNA extraction from buccal cavity

Cell collection

1. A taster and a non-taster of the compound phenylthiocarbamide (PTC) was identified by tasting papers dipped in PTC solution.

2. Cell suspension from both the taster and the non-taster was collected by mouthwash with 0.9% NaCl solution.

3. The cell suspension was centrifuged at 5000 rpm for 20 minutes. [In the first place, centrifuge needed to be carried out at 12000 rpm for 5 minutes but as the larger centrifuge had the max speed of only 5000 rpm, centrifuge was done for longer time]

4. Supernatant was discarded and the pellet was mixed with 2 ml 0.9% NaCl solution.

5. The suspension was centrifuged at 12000 rpm for 4 minutes.

6. Supernatant was discarded and the pellet was stored.

DNA extraction

• DNA extraction was carried out using Bioron Tissue DNA kit

DNA concentration

• Taster : 58.3 ng/µl [260:280 = 1.77]

• Non-taster: 25.7 ng/µl [260:280 = 1.74]