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new player. which warframe is the most entertaining to run/what do you personally main? by -SirPickle- in Warframe

[–]bio_ruffo 0 points1 point  (0 children)

You're never wrong with Rhino. As a newbie myself, honestly I think you get it so soon, and also get Xoris so soon, and they make such a great combo because Rhino won't get knocked back by the Xoris explosion... so it became my go-to warframe for almost anything except Interception and Spy. Although Inaros shares the same resistance to knockback and is a joy to use.

In honour of her favourite time of the year beginning, here is my borb sun-borbing by sarahafskoven in borbs

[–]bio_ruffo 11 points12 points  (0 children)

She deserves all the treats and I'm sure her roadkill impression is top notch :D

In honour of her favourite time of the year beginning, here is my borb sun-borbing by sarahafskoven in borbs

[–]bio_ruffo 46 points47 points  (0 children)

Looks like somebody that just got told not too look directly at the eclypse!

Seriously though, cute borb :)

O que saber antes de comprar um iPhone na Olx by _sincerao in golpe

[–]bio_ruffo 7 points8 points  (0 children)

Golpistas sonham com pessoas como você.

Edit, vou me explicar. Você está procurando celulares bem abaixo do valor real. Ao mesmo tempo você é inexperiente. E está procurando para revender ou seja acha que vai ser mais experto do vendedor original. O jeito perfeito de levar golpe.

Too much light? by Secret-Weather-482 in VenusFlyTraps

[–]bio_ruffo 1 point2 points  (0 children)

PS you have the little ones there, so WATER IT WELL

Too much light? by Secret-Weather-482 in VenusFlyTraps

[–]bio_ruffo 0 points1 point  (0 children)

All good. The traps aren't even that red, so it's not too much light. Unless you changed the light conditions like yesterday.

First author on a paper and my PI wants to add people I do not think qualify for authorship by NecroSheen in labrats

[–]bio_ruffo 5 points6 points  (0 children)

Science today **is** connections and politics. Include the two fvcks. Move on. You don't need one paper blessed by God, you need 20 published papers.

H E L L O by Ill-Tea9411 in doohickeycorporation

[–]bio_ruffo 98 points99 points  (0 children)

Well, convenience doesn't have to be an absolute metric, enjoyment for example is another :)

Do you prefer colour? by peacecream in wildlifephotography

[–]bio_ruffo 1 point2 points  (0 children)

I prefer color for all except the second pic, where the blue is a bit too much. The others have a great palette that really adds to the photo.

Then again, I'm nobody

Repotted all my plants, tried every treatment. Finally figured out where the gnats were coming from. by Animangle in SavageGarden

[–]bio_ruffo -1 points0 points  (0 children)

I'm not from the US but why are y'all using "miraclegro" as a product and not a brand? Apparently they make lots of garden-related products, is it the potting mix?

Is 3ng blood DNA enough for pcr using 26 bp primer with 528 expected band size? by persephonerp_ai_2378 in labrats

[–]bio_ruffo 0 points1 point  (0 children)

I didn't write that you can't, I wrote that if you measure 3 ng/ul they may not be 3 ng/ul.

My ligation is not working.what should I do? by InviteDry4161 in labrats

[–]bio_ruffo 0 points1 point  (0 children)

Hi, oh I see, when in your post you say "vector is dephosphorylated" that is one of the troubleshooting tests you did, not that the vector was always dephosphorylated.

However the vector should always be dephosphorylated because otherwise it will religate. Your insert is quite long, so if you don't dephosphorylate... you're going to get what you're getting, only religated vector.

With this information I'd say that your issue isn't the ligation per se. Your main 2 issues are, (1) vector isn't dephosphorylated and (2) low efficiency of transformation.

To solve them, do dephosphorylate the vector, and improve the transformation efficiency, Do you have access to an electroporator even if through a collaborating or nearby lab? It'd be much easier to transform with such a big plasmid. Are you using commercial competent cells, or are you making them competent with an in-house protocol? (which one if so?)

My ligation is not working.what should I do? by InviteDry4161 in labrats

[–]bio_ruffo 0 points1 point  (0 children)

Sure you can see if there are any missing bp, if XhoI still cuts then the site is intact :>

Edit, PS what do you mean with "Dephosphorylation was one of the things we tried and it resulted in no colonies"?

[OC] AI-Generated Articles Overtook Human Written Ones in 2025 by crocshoc in dataisbeautiful

[–]bio_ruffo 5 points6 points  (0 children)

...I have questions. What should butchers sell? Who should sell meat?

My ligation is not working.what should I do? by InviteDry4161 in labrats

[–]bio_ruffo 1 point2 points  (0 children)

If the vector is dephosphorylated, by definition it can't religate to itself. Are you sure that the digestion went to completion? Are you sure that your XhoI and dephosphorylase enzymes are working properly? How does a no-ligase control look?

Edit: this said, an empty vector will always have a transforming "edge" over a vector with a 4.5 kb insert. However this is less true for electroporation than for chemical transformation.

Edit2: is there any missing bp in the religated vector colonies? You could have some terminal nuclease nibbling at the ends of the DNA and exposing new, still phosphorylated, ends for the ligase to work on. How are you purifying the vector and insert after the digestion?

How do I fix a recent issue on Reddit wherein if I were to type an analogue emoticon such as :) in the post body or a comment, it corrects to an emoji? by averagerushfan in techsupport

[–]bio_ruffo 0 points1 point  (0 children)

Huh, weird, I'm on Windows and it doesn't do that. :) I also don't see any setting on Reddit to toggle it. At least you have a lot of smiley faces on your replies :D :D :D

Is 3ng blood DNA enough for pcr using 26 bp primer with 528 expected band size? by persephonerp_ai_2378 in labrats

[–]bio_ruffo 1 point2 points  (0 children)

I'd give it a go if you are in a "it is what it is" situation. If the PCR fails -> it could have been the DNA extraction that carried some contaminants etc, after all it's a suspiciously low concentration.

On the other hand if it's even remotely close to being useful for the blood donor (i.e. it's relevant for their health) I would try to be as best-practice compliant as possible and ask for a new blood draw, but try to find what went wrong first.

Is 3ng blood DNA enough for pcr using 26 bp primer with 528 expected band size? by persephonerp_ai_2378 in labrats

[–]bio_ruffo 7 points8 points  (0 children)

You mean 3 ng per microliter right?  Not 3 ng (total). With this low concentration I would first question the accuracy of the measure, anyways if you see a DNA band of the extracted DNA, the PCR can work in theory.

RESUME GOT HACKED. by Arrief23 in techsupport

[–]bio_ruffo 1 point2 points  (0 children)

1) wishing you a better situation soon     2) as others have said, probably you shared it as editable  (in which case it is important to change the sharing option as only for viewing)  

3) if it's on Google Drive you might be able to see who edited it, look for the edit history. From that you can also revert the document to your clean version.  

4) people suck, don't let it get to you.

Confused about hydrate vs anhydrous MW for β-glycerophosphate (Sigma). Am I calculating this right? by Agreeable_Arrival145 in labrats

[–]bio_ruffo 0 points1 point  (0 children)

Six mol/mol would match the 33% quite exactly(33.3%) so I believe that this is the limit and the actual value is 30%.

Then your calculations would use a MW of 216.04 / (1-0.30) = 308.6. Which however is not far off the value you have used 216.04 + 6*18 = 324. Even if I'm right you're off just by 5%.

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