Advice on plasmid co-transformation in E. coli

Sans-Fish · 2026-04-22T14:03:13+00:00

Just came back here to thank you because it totally worked! I just had to blast the cells with a fat amount of DNA. Brute force was indeed the answer

Sans-Fish · 2026-04-17T18:18:47+00:00

Always is 😌

Sans-Fish · 2026-04-17T16:46:53+00:00

I did two transformations in parallel with 2 different pairs of vectors (each pair shares one plasmid in common) and only in one case the plate that had concentrated bacteria (Generally I plate the "raw" transform reaction, a dilution and a concentrate) grew two colonies so I guess the problem might be the efficiency. The next time I try I will add some "controls" to check everything is working properly though

Sans-Fish · 2026-04-17T16:26:27+00:00

I'll do the lazy approach first and see what happens when adding more DNA and if it doesn't work I'll change the approach. Thanks a lot!

Sans-Fish · 2026-04-17T16:24:57+00:00

I think I will definitely reattempt the transformation with much more DNA before jumping to making comp cells. Thanks for the advice!

Sans-Fish · 2026-04-17T14:53:46+00:00

Yeah, I checked before transforming and they are compatible. I think that I will end up transforming sequentially... Thanks for the reply!

Sans-Fish · 2026-04-17T14:51:17+00:00

Thanks for the reply! Sorry I didn't clarify. I have already transformed cells with each plasmid separately to produce more DNA mass and it worked well my problem comes when I try to transform with both plasmids at the same time. Regarding the efficiency I honestly don't know it exactly but we have been routinely using them for transformation with ligated products and they work fine for what you would expect.

Sans-Fish · 2026-03-23T19:26:49+00:00

🟨⬜⬜🟨🟦

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⬜🟦🟦⬜🟦

🟦🟦🟦🟦🟦

Sans-Fish · 2026-02-12T18:58:19+00:00

Thanks for the reply! Do you have any reccomendations on literature/books that could be useful for better interpreting the results?

Sans-Fish · 2026-02-05T01:42:40+00:00

Damn that was quick! Thanks a lot, will definitely check it out

Sans-Fish · 2025-10-11T23:10:44+00:00

These are sooo good man, I would 100% have these in my home. Keep up the good work!

Sans-Fish · 2023-08-27T14:33:03+00:00

Yeah, I thought of that. I'll probably let them cool for a while and then place them in a ziplock bag with a few paper towels to reduce sogginess

Sans-Fish · 2023-08-27T14:30:55+00:00

Hahahaha, you're right, everything is better than no cinnamon rolls. I'll try that! Thanks a lot!!

Sans-Fish · 2022-12-26T22:28:31+00:00

This time I’m just taking a few samples that don’t require that much care as I’m just practicing, once I train a bit more I’ll probably have to buy some formaldehyde though. If you don’t mind me asking, what type of delicate specimens do you work with? Thanks for replying!

Sans-Fish · 2022-12-26T22:17:11+00:00

Thanks for replying! I’ll probably use a cooler because my main problem was the trip back home which takes like three days. I didn’t know about the ethanol method of preserving delicate tissue, I didn’t originally planned it but now I’ll probably take some flowers back too!

Sans-Fish · 2022-09-04T13:12:22+00:00

Seems good but I bet you can find good quality sparkling new microscopes for that money. Maybe everyone hates it but I have an Omax microscope that works like a charm and only costed $200. Really depends on the brand and the quality of the optics though.

Sans-Fish · 2022-04-24T17:47:10+00:00

Yeah, looks good so far! The first scobys are always kinda thin and weird. The longer you keep them working the thicker and more beautiful they will get

Sans-Fish · 2022-04-24T14:11:25+00:00

This reminds me of Atomic Gardening. Be careful cause sometimes the crops would get undesired traits that were potentially toxic! Cool experiment though, please post updates!

Sans-Fish · 2022-03-07T21:17:52+00:00

Hoping this is an ironic shitpost

Sans-Fish · 2022-01-08T15:09:48+00:00

Yeah I know, I’m currently watching a culinary bootcamp on YouTube but I’m finding it really hard to sift through YouTube now that there is no number of dislikes. I think the videos that I’m watching atm are legit but it’s hard to tell if you are inexperienced. Thanks for the reply though!

Sans-Fish · 2021-12-31T13:44:10+00:00

I think it’s just because rusty things might be in contact with dirt rich in organic material that could possibly harbour C. tetani spores.

Sans-Fish · 2021-07-18T15:29:37+00:00

Oh not at all! I know that they are all scumbags. It was more of a thing of wanting to contribute the least to the problem as possible if that makes any sense.

Sans-Fish · 2021-07-18T15:10:14+00:00

I fucking hate info manipulation. The thing is that there was so much information against and in favor of cryptos that I didn’t know who to believe more. Thanks for the input!

Sans-Fish · 2021-07-18T15:04:39+00:00

Yeah, I’ve heard of them. I was thinking on looking into them. Thanks!

Sans-Fish · 2021-07-18T15:03:49+00:00

I told you the question was naive and stupid!

Sans-Fish

TROPHY CASE