Re: [DIYbio] Re: stable transformation of chlorella vulgaris

Also, I've been told that maintaining a "pure" culture of algae can be extraordinarily difficult and you may want to include some sort of permanent selection factor (i.e. they need the plasmid to survive so they will continue to retain it over long periods of time).

-SG

On Wed, Jul 11, 2018 at 5:51 PM, Skyler Gordon <skgor1@gmail.com> wrote:
Chances are that when you go to isolate the genetic material and look for your insertion site, there will be plenty of remaining vector. I'm not sure what method you were going to use to determine this (qPCR, gel validation, sequencing, ddPCR, etc.) but here is an interesting paper about how Eukaryotic cells retain their vectors for quite a while.


I hope this helps

-SG

On Wed, Jul 11, 2018 at 12:01 PM, Scott <synbiofablab@gmail.com> wrote:
Hello Najib,

This reference might help you with designing your electroporation experiment. You will need to linearize your vector for stable integration.

There are many protocols for looking at transgene integration sites, inverse PCR being the most popular. There are even kits but I'm old school.

Compare your integration sequences with the Chlorella vulgaris assembly.

Hope this helps and all the best with your experiments.

Cheers,
Scott

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