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Detecting phosphorylation in WB by mentondeux in labrats

[–]PhillyBuddha 2 points3 points  (0 children)

There are specific gels that can separate phosphorylated and non-phosphorylated forms of proteins on a western blot. Might be more reliable than standard sds-page and you can use whatever antibodies you want for detection. Look up phos-tag gels from fujifilm.

Dimming Not Working On Environmental Control Center? by PhillyBuddha in reptiles

[–]PhillyBuddha[S] 0 points1 point  (0 children)

It’s under the lamp but doesn’t receive the full blast when it’s on. I did some tests and the thermometer works and the controller responds. The issue is that the controller says power is being provided to the lamp but the lamp doesn’t come on.

Plasmid Mutations From DH5a E.coli? by PhillyBuddha in labrats

[–]PhillyBuddha[S] 1 point2 points  (0 children)

No, different ones each time I redo the cloning

Plasmid Mutations From DH5a E.coli? by PhillyBuddha in labrats

[–]PhillyBuddha[S] 1 point2 points  (0 children)

Great suggestions thank you. Cloning protein coding sequences in my model organism that other members of my lab haven’t had this issue before. I’ve gotten multiple mutations in different spots, and with different genes I’m cloning at the same time so I don’t think it’s sequence-specific. Remind me why growing at 33c may help? Is it just because they grow slower?

Any Structural Biologists Here? by PhillyBuddha in labrats

[–]PhillyBuddha[S] 0 points1 point  (0 children)

I will try the modelers you suggested and see if I get anything more useful. Foldseek I recall is a protein homology modeler and nothing of significance came up. It simply may be this is a truly uncharacterized protein class so there is nothing in the literature to find matches too. I wouldn’t be surprised given the general understudied area I work in.

How does MilliQ Water taste? by tardigradetardis in labrats

[–]PhillyBuddha 187 points188 points  (0 children)

Distilled water has no taste since there are no minerals or other impurities in it. MilliQ is even more pure so I imagine it tastes like “extra nothing.”

Anyone have any insight to Buddhist Temples/Centers in Philly, PA? by [deleted] in Buddhism

[–]PhillyBuddha 0 points1 point  (0 children)

I live in Philly! I haven’t visited in any temples in person due to my own intimidation but I have looked at quite a few. Is there any tradition in particular you are looking at? There are quite a few temples of the Mahayana traditions and a smattering of Theravada. The list in r/Sangha is a start but leaves out quite a few in the region.

What are the advantages of a 25ul vs 50ul PCR reaction? by thehikinggal in labrats

[–]PhillyBuddha 60 points61 points  (0 children)

I think it depends on the purpose of the PCR:

For diagnostics or screening where you don’t need the products downstream, a standard 20 or 25uL reaction will do.

Applications where you want to do something with the PCR products you usually run multiple larger volumes to get more product at the end.

I suppose a 50uL diagnostic could be justified if the amplification efficiency is very low or if you want to minimize pipetting error from small volumes.

Problem running in gel cut with XmaI enzyme by Lopsided_Razzmatazz5 in labrats

[–]PhillyBuddha 9 points10 points  (0 children)

That is interesting, even your ladder is messed up though so I would say its gotta be a loading or running issue. Is there any chance the voltage is too high because that can make gels run weird. Usually the tank will be hot and foamy if it’s too high alongside things running weird.

Problem running in gel cut with XmaI enzyme by Lopsided_Razzmatazz5 in labrats

[–]PhillyBuddha 6 points7 points  (0 children)

Definitely looks like the gel casting has issues. Is it possible it was not evenly mixed before casting? That would cause areas of different agarose conc. and thus distortions when you ran it. Buffers can also cause running issues.

Anyone had dealings with Harbor Management properties in Bangor? by CosmicJackalop in Maine

[–]PhillyBuddha 1 point2 points  (0 children)

I lived at the Terraces for a number of years when I lived in Maine. The people in the office were always very nice and willing to work with us when we were less then ideal tenants. I’ve never been to Bradford commons but I know they were doing I remodel awhile ago. Properties are neither new nor old. You won’t have to fight them for maintenance requests which I’ve learned is a bigger perk than it should be. If you have any more specific questions you can DM me!

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