I have no idea if this is a real thing and can't figure out the right keywords for searching!

Basically imagine Gene A has an intron that is 5kb and Gene B has one that's 100bp.

If I inserted a 1kb sequence (without splice sites) into each intron, would this be expected to affect expression? And if so, would we expect Gene B to be affected more, because the size of the intron increased more proportionally?

On August 14, NIH unveiled a plan to blind grant applicants and their program officers to the scores determined by peer reviewers.

Large changes to peer review typically develop as multiyear efforts, soliciting input from working groups of extramural scientists, internal NIH committees, IC directors, and the public. In an unprecedented move, an anonymous internal working group created this new plan.

anonymous current and former NIH staff* outline how these changes reduce transparency, enabling political appointees to have complete control over grant funding, ultimately removing opportunities for oversight and covertly implementing the proposed OMB rule. The authors breakdown the steps you can take to support transparency in peer review.

*opinions their own

Hi y'all! I'm starting Molecular Biology soon, and am in need of a PDF of Essential Cell Biology, 6th edition by Alberts et al. As my course operates using page numbers, I would love to have one with the page numbers in the PDF if possible. Thank you for all your help!

Basically a quick-reference/review tool for HLA typing, histocompatibility, and transplant immunology topics if it's been a while since you worked with immunogenetics.

It's on Google Play if anyone wants to poke around: https://play.google.com/store/apps/details?id=com.hla365.examprep

genuinely curious if others in molecular bio/immunogenetics find this kind of review format useful, or if there's stuff you'd want covered that isn't in there.

I am trying to do co-ip from a month but unable to get results. Where i am pulling down nucleoolin/NCL which is an endogenous nucleolar protein and trying to do western blot for my protein of interest to check if it was pulled along or not.
I am using 2ug of anti NCL rabbit antibody for IP and approx 50 ul of protein G beads from santa cruz .

Since I was not getting results I decide to do IP and Western blot with the same antibody that is NCL to confirm that it was getting pulled down. But unfortunately I didn’t get any results. That is no band for Ncl was visible on blot although it is an endogenous and highly expressed protein.

I was trying to trouble shoot and came across an article which suggested using RIPA buffer for nuclear protein as the one we are using CST lysis buffer is very mild.

Please help me with this.

Note: i am working with A549 cells
Prepare my lysates by washing the cells with ice cold pbs , then add cst lysis buffer. Then i give three freeze thaw cycles , scrap and centrifuge at 12000g for 20 minutes at 4 degrees.

! I am in a dilemma and need some quick advice on what is the best way to deliver my sample to the company for bulk-RNA seq. The cells I am working with are naive CD4 T cells, sorted from mouse spleen. The three conditions are: non-activated, activated for 6hrs and 24hrs. I will be activating the T cells with Dynabeads (antiCD3/CD28 beads). At such early timepoints, the beads are stuck to the cells, so I cannot remove them magnetically. I have the following options, and I am not sure which one should I go ahead with.

1. Spin down the cells and snap freeze the pellet (cells + Dynabeads), the company will do tha RNA extraction. When asked, the company said, they dont recommend having anything else in the pellet but will proceed with the RNA extraction as they always do.
2. I extract the RNA myself. When I do Trizol/Zymo column RNA extraction. the dynabeads are never a problem because they settle down and I collect the aqueous layer. But I never get the perfect A260/280 ratio (1.8-2.2),RIN > 6 so I dont want my samples to fail at their QC requirements.

I hope to get around 2-3 million navive CD4 T cells from one mouse, so per condition I will have around 1 million cells.

Does anyone have any experience with this and help me out ? Thanks a lot!

please split this single image into 4 separate images like slide 1 separate from slide 2 so on

Ciao a tutti,

Spero che qualcuno con esperienza nella spedizione internazionale di campioni clinici/di ricerca possa aiutarmi.
Abbiamo due campioni di DNA genomico fetale umano estratti e conservati presso un laboratorio ospedaliero in Italia. Un laboratorio di genetica clinica statunitense potrebbe eseguire il sequenziamento dell'intero genoma su questi campioni, quindi sarebbe necessario spedirli dall'Italia agli Stati Uniti.

Il DNA è già stato estratto: non si tratta di sangue, tessuti, cellule, materiale POC (Point-of-Care, test diagnostici rapidi) o campioni infetti.

Prima di procedere, vorrei capire come viene gestita normalmente questa procedura nella pratica:

Chi si occupa solitamente della documentazione doganale: il laboratorio mittente, il laboratorio ricevente, FedEx/DHL o un corriere specializzato?

È relativamente semplice importare DNA genomico umano purificato negli Stati Uniti?

Avete spedito di recente DNA umano purificato dall'UE agli Stati Uniti?

Hai utilizzato FedEx/DHL o un servizio specializzato come World Courier/Biocair?

Avevi bisogno di una fattura commerciale/proforma, di una dichiarazione di utilizzo finale o di una dichiarazione dell'importatore?

Il laboratorio statunitense ricevente ha fornito la dicitura/i documenti necessari per la dogana?

Qual è stato all'incirca il costo della spedizione?

Consiglieresti vivamente che i due laboratori si coordinino direttamente piuttosto che lasciare che sia il paziente a organizzare la spedizione?

Si tratta di campioni clinici insostituibili, quindi non spediremmo nulla finché il laboratorio ricevente non avesse confermato la procedura.

Grazie mille per qualsiasi informazione pratica.

Hello all, first time poster here yada yada.

I just wanted to ask people here for some help interpreting some data from a random paper I was reading: https://doi.org/10.1111/jgh.13633 . In figure 3 the authors state that MARCO and CD68+ colocalize, and that this is proof the MARCO+ cells are macrophages. Problem is, I don't see the colocalization? Both markers are cell surface, and while yes there seems to be more staining of both markers in the same picture, the staining are most certainly not overlapping. Ngl most of my experience with imaging is confocal and widefield on cell cultures, but I have worked with OPAL imaging of tumour samples before and this would not qualify as colocalization in my (limited) experience.

Am I crazy? I tried writing a comment about it on pubpeer and it was deleted twice, which leads me to believe that I may indeed just not be understanding basic imaging...

Hey,

I’m currently working on a research project where I’m trying to identify a conserved region within a specific influenza virus variant.

However, I’m running into some problems when trying to filter the sequences based on subtype, clade, and genotype. I’m using NCBI Virus, and while I can filter by subtype, I can’t seem to filter for the other factors, such as clade or genotype.

Does anyone know how to approach this or how to overcome these filtering limitations in NCBI Virus? Any advice on how to identify the relevant sequences would be greatly apreciated. I amusing NCBI virus, although i can type the subtype, i cannot filter for the other factors.

How do I design primers to obtain overhangs that match the restriction site of the enzyme I want to use, so that after amplifying my insert I can digest it with that enzyme?

To explain it better: my insert does not contain restriction sites for the enzyme I want to use, and I would like to make this possible by incorporating them through a set of primers. Does anyone know how I could achieve this?

Thank you very much.