Hi Kanwar, after the PCR, we check if the PCR was successful by running the reaction mixture in a gel (gel electrophoresis). Then, we treat the successful reaction mixture with some enzyme before we send them for sequencing to any sequencing services. you can check the following publication in the material and method section. www.exeley.com/journal_of_nematology/doi/10.21307/jofnem-2020-098
Could you please tell me, what you have done after thermocycler steps? How many cycle do you have considered? Here you didn’t use any master mix or other ingredients in pcr mix. I would be very greatfull if you could tell me the next steps.
Hi Mr. Khan, This video demonstrates only upto DNA extraction step. The extracted DNA can be used for further PCR. You can use any commercially available PCR master mix and your desired primers for PCR according to your needs. Master mix and PCR cycles or thermal profile can vary from person to person, also according to targeted gene etc. Good luck
I suggest you register for the Nematology Digital Learning Platform: www.imanema.ugent.be/workshops/learningplatform Here you can find the protocol and much more!
Can you please tell what you have done after PCR step. Do you directly done the electrophoresis or some additional steps are there..??? Please tell me
Hi Kanwar, after the PCR, we check if the PCR was successful by running the reaction mixture in a gel (gel electrophoresis). Then, we treat the successful reaction mixture with some enzyme before we send them for sequencing to any sequencing services.
you can check the following publication in the material and method section.
www.exeley.com/journal_of_nematology/doi/10.21307/jofnem-2020-098
Could you please tell me, what you have done after thermocycler steps? How many cycle do you have considered? Here you didn’t use any master mix or other ingredients in pcr mix. I would be very greatfull if you could tell me the next steps.
Hi Mr. Khan, This video demonstrates only upto DNA extraction step. The extracted DNA can be used for further PCR. You can use any commercially available PCR master mix and your desired primers for PCR according to your needs. Master mix and PCR cycles or thermal profile can vary from person to person, also according to targeted gene etc. Good luck
Can you please share the protocol/procedure used to prepare the worm lysis buffer. I have been struggling with isolating DNA from a single worm.
I suggest you register for the Nematology Digital Learning Platform: www.imanema.ugent.be/workshops/learningplatform Here you can find the protocol and much more!
Hi Glen, You can also now find a new video in the channel on how to prepare the worm lysis buffer. Good luck.
Hello@@imanemaugent9505 , I can not visit this website to find the protocol. Could you please help? Thank you.