User:Saroj Pandey/Notebook/SNP PCR optimization/2014/09/17: Difference between revisions
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'''Cell collection''' | '''Cell collection''' | ||
1. A taster and a non-taster of the compound phenylthiocarbamide (PTC) was identified by tasting papers dipped in PTC solution. | 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. | 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] | |||
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. | 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. | 5. The suspension was centrifuged at 12000 rpm for 4 minutes. | ||
6. Supernatant was discarded and the pellet was stored. | 6. Supernatant was discarded and the pellet was stored. | ||
'''DNA extraction''' | '''DNA extraction''' | ||
• DNA extraction was carried out using Bioron Tissue DNA kit | • DNA extraction was carried out using Bioron Tissue DNA kit | ||
'''DNA concentration''' | '''DNA concentration''' | ||
• Taster : 58.3 ng/µl [260:280 = 1.77] | • Taster : 58.3 ng/µl [260:280 = 1.77] | ||
• Non-taster: 25.7 ng/µl [260:280 = 1.74] | • Non-taster: 25.7 ng/µl [260:280 = 1.74] | ||
Revision as of 12:44, 21 September 2014
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gDNA extraction from buccal cavityCell 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] | |
