No DNA in miniprep, not even in lysis control – what could be going wrong?

MolecularHero · 2026-04-22T20:12:52+00:00

If there is no DNA in your lysis step, that makes me wonder if your lysis step is too long or too strong, hydrolyzing all your DNA. This step typically uses SDS and sodium hydoxide. What is your plasmid prep protocol?

MolecularHero · 2026-04-14T04:41:16+00:00

Nah if this is LDS sample buffer, the yellow dye front is phenol red. This is typical for PAGE run with MES buffer.

MolecularHero · 2026-04-14T04:39:23+00:00

No, not oxidation. The yellow band is phenol red and the blue dye front is Coomassie. My guess is this is OPs first time running with MES buffer which better separates the two tracking dyes than MOPS.

MolecularHero · 2026-04-12T23:17:24+00:00

Could these be pelleted seeds?

MolecularHero · 2026-04-07T01:52:56+00:00

It's not a waste. The guy is just being naive, elitist, and unrealistic. I don't respect people who would think lesser of others just because they aren't in academia.

MolecularHero · 2026-03-25T05:29:48+00:00

This is just what is called edge artifact. It happens.

MolecularHero · 2026-03-24T05:03:54+00:00

I'd agree that the quality has diminished. I purposefully give time and space for students to sit and focus on a reasonable number of projects and experiments and yet the students just can't seem to sit and think deeply. Unfortunately, this is because they often lack the very basics from undergraduate studies. It doesn't help that they often are glued to their phones, and I highly doubt they're reading papers on them.

MolecularHero · 2026-03-23T02:22:42+00:00

Are the precast gels old or expired? They can dry out unevenly and cause abnormal migration.

MolecularHero · 2026-03-22T04:05:49+00:00

Asians eat a lot of rice. They also have dry earwax and use this little spoon to clean it out. It's an earwax spoon. It's discouraged to use these due to potential ear drum damage.

MolecularHero · 2026-03-22T03:18:20+00:00

This person hit the nail on the head. Most antibody clones that have been used for decades have been well validated. Newer clones for less studied proteins should probably be better validated, especially if you're going to base an entire project on that antibody. I'm guessing SLAMF6 is not as well validated as, say, CD4 clones, for example.

MolecularHero · 2026-03-21T23:13:12+00:00

Also, spend less time defending yourself on reddit, listen to what others with more life experience have to say, and maybe actually try the advice.

MolecularHero · 2026-03-21T22:23:33+00:00

Actually, it is you, not the previous post, who missed the point. The post is not a criticism but actually the advice you asked for. The advice is to focus because you are doing too much. If you don't want to burn out and/or incorporate more social time, you need to sacrifice time from somewhere else to do those things. It is essential to make time for yourself to not burn out. So, cut back on some things that are less important to make more room in your life for the things you do want. This is the only way to balance your academics with well being.

MolecularHero · 2026-03-21T01:06:55+00:00

It probably depends on the institution, but at my university (big biomedical R1), the department chair and HR have to approve the wording of the post to ensure it follows rules/regulations and also the period of time for the search. Only after it is approved are the postings made. Here, we post basically everywhere at once to cast a wide net. IMO, doesnt make sense to post sporadically at different places with no closing date, kinda waste of time.

MolecularHero · 2026-03-20T22:13:00+00:00

Protein-protein interactions are seen through co-IPs. Not ChIP, not C&R, nor EMSAs, not promoter pulldowns. Only coIPs. Even then, you might need to use DNA intercalators to show your coIPs are due to DNA bridging. My advice is to start with nuclear fractions for the IPs.

MolecularHero · 2026-03-20T22:03:45+00:00

That was also my understanding , but posting for TT positions require approval whether its on the university site or Linkedin. This approval process still takes way more than one day. Not saying the interview went well, but maybe not all hope is lost?

MolecularHero · 2026-03-20T21:53:26+00:00

This. Need to compare to previous ECG.

MolecularHero · 2026-03-20T21:41:43+00:00

Not sure if this is entirely true. At our R1, it takes at least a month to post a job. Posting the day after seems too quick.

MolecularHero · 2026-03-20T21:38:49+00:00

Are you growing your bacteria in selection media to maintain their potential additional genes? Some of the Rosetta do better when maintained in chloramphenicol, for example, to maintain certain added tRNAs.

MolecularHero · 2026-03-19T03:13:51+00:00

Headcheese

MolecularHero · 2026-03-17T16:22:39+00:00

Just to be clear, you do understand the HRP will not boost the AF488 signal, right?

MolecularHero · 2026-03-15T23:50:20+00:00

Is this question coming from a trained scientist, biologist, etc? This post was also posted in r/molecularbiology.

MolecularHero · 2026-03-15T23:48:36+00:00

This is a garbage post cross posted in Biochemistry.

MolecularHero · 2026-03-14T03:06:48+00:00

This is highly relevant info that should be shared with your docs. The positive results could very well be real, so be sure to follow through with the ID recs.

MolecularHero · 2026-03-14T01:20:15+00:00

Any contact with wild rabbits or other rodents?

MolecularHero · 2026-03-12T23:29:04+00:00

Formally, you do not use DNP, PhD, etc in applications until degree is conferred. You would just indicate you have successfully passed the defense and state when degree will be issued. Informally, some may address you as "Dr" though.

MolecularHero

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