I used a protocol mentioned below and it did not work.
dissolved single colony with 100ul of buffer(200mM LiAc,100mM SDS)
incubated at 70C for 5min
Added 300ul of icecold ethanol and vortexed briefly.
incubated at -20C for 1hour.
centrifuged at 15000xg for 3min
washed the pellet with 70%ethanol(200ul)
centrifuged at 15000xg for 2min
air dried the pellet and dissolved in 100ul of molecular grade water
centrifuged at 13000xg for 1min
and used 2ul of supernatant as template for PCR amplification.
Are you sure that your PCR/primers are working? Did you use a positive control?
several links on researchgate where you may find valuable answers :
https://www.researchgate.net/post/Does_anyone_know_how_to_do_a_PCR_reaction_from_a_single_yeast_colony
https://www.researchgate.net/publication/235419692_Comparison_of_colony_PCR_methods_for_yeast
https://www.researchgate.net/topic/colony_pcr2
Conference Paper Comparison of colony PCR methods for yeast
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