I set up a viral enrichment by inoculating 5 ml of Host culture in late exponential stage with 1 ml of filtered water (viral sample) for 1 week. The solution cleared up and I centrifuged the cleared culture and filtered the supernatant. Then I made serial dilutions 10-1 to 10-5 and performed plaque assay using overlay method [150 microlitre host culture and 100 microlitre viral sample enriched]. However I am not getting any plaques on the plates. On some of the plates the bacterial lawn have formed. What should I do?
Solid agar concentration 1.5%
Overlay agar concentration 0.5%
Media = high salt 200g/l
pH = 4.0
Before doing agar overlay , do spot test with filtrate
Then check your bacterial culture whether it got contamination, spot test for viral confirmation, streaking for bacterial purification , if both are alive and pure form, definitely u got good results ,
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