Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Plant research: Watering uniformity/consistency. How important is it?

Trying to gather insight from an unresolved discussion with a colleague here, and either the available literature surrounding this topic and/or my keywords in search of are lacking. Context:...

21 February 2021 6,389 3 View

How to get rough estimation of the concentrations of Oxygen and moisture inside the glove box?

Dear colleagues, I was just wondering if you can suggest a good method to get a rough estimation to the concentration of O2 and H2O inside the glove box as unfortunately, our glove box doesn't...

17 February 2021 1,012 3 View

Onset of Photo Oxidation in Polymers?

Hi! I am looking for some specific information in regards to the minimum amount of UV intensity required to begin the photo degradation process in polymers. I understand each polymer is...

15 February 2021 7,703 4 View

Are you looking for a quality online conference on teaching and learning technology?

This large international conference will be held online this year due to COVID. I am submitting a paper to the Technology Enhanced Language Learning track (8). It is about a topic I have never...

15 February 2021 1,975 15 View

How do you calculate a discrimination index for polytomous data?

Hello, I'm currently running a PCA on a polytomous data set (5 point Likert). I want to create a discrimination index to narrow down my items before running the PCA. After looking around, I'm...

13 February 2021 3,517 2 View

Regarding immunology, transmissivity, herd immunity and vaccines in Sars CoV-2?

This is a follow-up on an earlier question regarding mechanistics of vaccines: why current vaccines don't block transmission. In summary, they don't engage mucosal immune cells to produce IgA....

08 February 2021 8,321 3 View

Which tropical country is the most easy for obtaining research permit (for biological research)?

Dear all, High beurocracy hihly complicate field ecolocial and evolutionary research nowadays. Nearly all tropical countries (maybe all?) requires research permit for performitng research and in...

04 February 2021 3,672 6 View

Can I polymerize NIPPAm with PEGmethacrylate or only with PEGacrylate?

I want to perform the synthesis of a hydrogel and I want to know if I can polymerize NIPPAm with PEGmethacrylate or only with PEGacrylate?

02 February 2021 2,106 6 View

What is the maximum storage time for samples used in soil enzyme assays?

01 February 2021 4,616 3 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

What urea buffer can i use for biotagged protein extraction from streptavidin beads?

Hi all, I have been doing research on biotagged LDB1 proteins in Mel cells. I purify the proteins using M280 streptavidin beads. The yield is very low so I've been trying to optimize it by...

24 February 2021 5,749 3 View

Can I use a protein-devoid condition as my negative control in ELISA?

Hello, I am running a sandwich-type ELISA where I am trying to inhibit a protein-protein interaction using small molecules. For each experiment, I determine my Z' (Z-prime) to determine the...

23 February 2021 7,429 1 View

Is there a way to extract total RNA and protein from cell culture at the same time, without denaturing protein?

Hi all, Is there a way to extract RNA and protein from the same cell culture sample? Because we need to use the protein for gelatin zymography at later time, we are looking for a method that can...

22 February 2021 5,341 5 View