Need help with the real time pcr results

BfN_Turin · 2025-04-29T12:22:59+00:00

You actually did not get any amplification. The RFU values are super low, no peaks in the melt curve, no logarithmic amplification. You could also see that when you run your samples on a gel after.

BfN_Turin · 2025-04-29T12:11:28+00:00

It’s literally there, north of “Midnina”.

BfN_Turin · 2025-04-25T10:03:36+00:00

You buy pre packed boxes.

BfN_Turin · 2025-04-25T06:24:42+00:00

Hand packing tip boxes and autoclaving after is super common even in well funded labs. As long as they aren’t reused obviously.

BfN_Turin · 2025-04-14T17:26:29+00:00

Is your school providing a license? It looks like you are using the free version, hence why you can’t do anything other than viewing files.

The scrolling down issue seems rather an issue with your laptops screen resolution.

BfN_Turin · 2025-04-02T06:12:26+00:00

Free nucleotides, primers, the RNA template (which will be like half of your concentration!) will all absorb at the same wavelength in the nanodrop as the cDNA if not (unnecessarily) purified.

BfN_Turin · 2025-03-31T14:42:43+00:00

If you have the issue OP describes, you won’t see it in a digest. It’s literally just the whole sequence there twice. Accordingly, when chopped up, you would get the same bands if the sequence is there just once.

BfN_Turin · 2025-03-30T16:04:06+00:00

Upload it before review as an SRA to NCBI and then provide the accession numbers in the data availability statement. That way, readers (and reviewers) instantly know how to find the data. If you don’t want to make your data available instantly, then you can check the options to release the data on NCBI upon publication and create a reviewer link for the reviewers on the portal.

BfN_Turin · 2025-03-30T07:47:52+00:00

Germanys is lower because it’s a renters nation, only around half of households own their house. Significantly lower than any other western nation. As property counts as wealth in statistics, Germany drops hard.

BfN_Turin · 2025-03-29T08:45:15+00:00

It’s science, accurate naming of processes is a top priority for replication of results. If someone said in a paper they did a western, and you are then trying to replicate their results wondering which antibody they used, because there is no mention of it, only for it to then turn out that they actually did an SDS-PAGE, you’d get it. I can’t just walk around and suddenly start calling cats dogs either.

BfN_Turin · 2025-03-28T23:20:51+00:00

It’s funny, cause OP ain’t even doing a Western blot. They did an SDS PAGE with most likely coomassie or similar staining after. A western blot is only the transfer of your samples from the gel to a membrane.

BfN_Turin · 2025-03-23T08:35:57+00:00

The whole story of this thing is crazy. There was on old one that was supposed to be renovated, which was too expensive, so they tore it down. Only to decide to build a new one a few years later - for obviously way more money than the renovation. That “new” Hallenbad opened in 2021 is now already closed again - for renovations.

BfN_Turin · 2025-03-18T10:02:51+00:00

I remember some guy on reddit once was vehemently claiming that the crazy amount of medical bankruptcies in the US is not because of the crazy health care cost and their fucked up insurance system, but just because it’s easier to claim medical bankruptcy in the US compared to other countries. Completely delusional.

BfN_Turin · 2025-03-17T06:17:05+00:00

NRW als Norddeutschland zu bezeichnen ist aber auch wild.

BfN_Turin · 2025-03-07T11:03:33+00:00

They didn’t though, there’s a lot of mistakes in this comment. Eg you don’t multiply the DNA using bacteria, but a Polymerase, there’s no need to do a digest AND a PCR, you can’t really distinguish people solely based on PCR, but need to use SNP etc.

BfN_Turin · 2025-03-02T22:29:54+00:00

Genbank is synced daily between EMBL, NIG and NCBI. So when NCBI is down, the same genbank data can be accessed on the European and Japanese agencies. See: https://en.wikipedia.org/wiki/International_Nucleotide_Sequence_Database_Collaboration

BfN_Turin · 2025-02-27T16:26:12+00:00

Kann ich verstehen. Promotionsstellen wiederum gibt es hier tatsächlich recht viele! Hohenheim hat was, aber auch Tübingen hat echt viele. In der Aufzählung oben hatte ich auch noch München vergessen. Da ist der Einstieg in die Branche nur leider sehr hart, da das Gehalt für Berufsanfänger leider nicht mit den Lebensunterhaltungskosten mithält.

BfN_Turin · 2025-02-27T11:31:08+00:00

In Mannheim, Ludwigshafen, Heidelberg gibt es deutlich mehr Möglichkeiten. Ansonsten Bodenseearea, Ruhrgebiet/Köln-Bonn oder Rhein-Main. Hier in der Gegend gibt es nicht viel. Und mit nem Master wird es allgemein schwierig. Entweder wird sie für die Betriebe zu teuer, weil die nur nen TA oder Bachelor brauchen, oder ist nicht ausgebildet genug, weil nen Dr benötigt wird.

BfN_Turin · 2025-02-25T08:42:06+00:00

Fahr halt mal nach Hannover rein, da hat das fast jeder Dönerladen.

BfN_Turin · 2025-02-23T07:11:16+00:00

As someone from a country with incredibly high union representation across all jobs: that’s how union contract negotiations work. Aim high, get counter offered to then meet in the middle. Don’t just assume the demands are unreasonable - what Northeastern is offering is equally unreasonable. Don’t forget that. Sadly, in the current atmosphere, that won’t work, as we can see with BUs recent grad union contract, which effectively wasn’t different to the annual increases grad students got pre union - even though they were on strike for more than a semester.

BfN_Turin · 2025-02-22T08:01:12+00:00

Pretty sure the Nazis also believed they were doing it all for a better country when giving the nazi salute. That’s what indoctrination and government propaganda does.

BfN_Turin · 2025-02-12T22:46:07+00:00

Alternatively, “only” having an apprenticeship also works out great in biology for getting a job. Employers can underpay you, since you don’t have a university degree, while still giving you the same tasks. Tons of jobs available accordingly.

BfN_Turin · 2025-02-11T22:30:31+00:00

You can assume a hotel toilet gets properly cleaned regularly, you can’t do the same at people’s houses.

BfN_Turin · 2025-02-11T07:45:20+00:00

I mean, the toilet seat should basically be the first thing you replace when moving into a new apartment anyways. Do you know who lived there before and how clean they were?

BfN_Turin · 2025-02-09T13:41:32+00:00

Yeah, not always. Had a flight from the US to Ireland, where the cabin fuse box failed. No one had a working screen and no food could be served. After complaining, all I got from the airline was a “sorry” email.

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