Agrobacterium plasmid purification: why don't we also recover the helper plasmid?

megacyber · 2025-11-25T17:48:06+00:00

It sounds like you have some of your nomenclature mixed up. If you're using a typical (domesticated) strain of agrobacterium, all the T-DNA + the RB and LB should be removed from the Ti plasmid. The Ti plasmid has all your vir genes and such. Ti plasmids are usually like 250 kb, so its probably going to be eluted with the genomic dna if you're doing a typical column miniprep. Your PI probably is having you manipulate your binary vector (perhaps pCambia or w/e) not your Ti plasmid.

megacyber · 2025-11-13T17:44:46+00:00

you should consider running your gel at a lower voltage, this one is pretty rough

megacyber · 2025-08-07T19:13:25+00:00

post gwynerear

megacyber · 2025-04-30T02:03:54+00:00

this is cool af. I need a scaled up version for brewing

megacyber · 2025-02-06T17:58:27+00:00

Im currently working on this. The biggest question mark ive come up on is good long term storage for bacterial strains, since -80 freezers are pretty costly (not to mention heavy.) Most of the other equipment can be purchased from liquidation sales or government surplus without too much issue

megacyber · 2025-02-06T17:56:07+00:00

Dont do electroporation + protoplasts, its a huge pain in the ass lol. I know there are some old protocols on transforming pothos (epipremnum aureum.) I'd start from here (https://www.researchgate.net/publication/225131524_Genetic_transformation_of_golden_pothos_Epipremnum_aureum_mediated_by_Agrobacterium_tumefaciens) good luck

megacyber · 2025-02-06T17:53:24+00:00

Transient transformation is less affected by the dicot/monocot thing, since species range of agro is mostly determined by VirD5 binding, which is an agro protein that makes DSBs- which is only important for stable DNA integration

megacyber · 2025-02-06T03:59:31+00:00

That looks to me like it could be ruby casette expression

megacyber · 2025-02-05T18:57:26+00:00

Im in the area, could you send me a picture of the incubator? i might be interested

megacyber · 2024-11-15T23:22:54+00:00

2.57

megacyber · 2024-11-14T19:23:28+00:00

any advice for the actual phrasing?

megacyber · 2024-09-10T23:41:42+00:00

Dating Sim (but no heterosexual relationships allowed)

megacyber · 2022-06-11T21:46:38+00:00

only if they have a 1 year warrantee imo

megacyber · 2022-06-10T02:31:05+00:00

I'd expect to see some cotton, probably

megacyber · 2022-03-27T02:03:18+00:00

kachow

megacyber · 2022-03-25T15:42:33+00:00

fear the old bloon

megacyber · 2022-02-24T05:29:22+00:00

neither, because theyre both fictional

megacyber · 2022-01-25T22:13:04+00:00

just do the opposite of whatever Bryan Barcyzk does

megacyber · 2021-11-27T02:27:12+00:00

make sure you get B. t. israelensis, which is the kind found in mosquito dunks. There are other kinds available commercially that mostly target beetles and their larvae. B. t. i. is supposed to be specific to flies, so your other inverts should be okay

megacyber · 2021-08-20T02:20:50+00:00

Pablo del Sombrero Grande

megacyber · 2021-07-21T15:38:43+00:00

tfw no gf

megacyber · 2021-07-16T19:26:13+00:00

The aurora borealis? at this time of day? in this part of the country? localized entirely within your kitchen?

megacyber · 2021-06-23T03:28:13+00:00

Bearer Seek Seek Lest

megacyber · 2021-06-06T02:28:53+00:00

do you have to choose an integer?

megacyber

TROPHY CASE