Does anyone know of a more affordable alternative to Mito-Tracker, that is well retained in fixed cells?

I have in the past worked with the MitoTracker dies. They are rather expensive and that is the bit I am hoping to avoid. I would really appreciate it if anybody can suggest a mitochondrial...

05 November 2020 5,701 1 View

What is a good ligation strategy with compatible ends EcoRI and MfeI in the vector DNA and the insert?

I used to do this kind of thing and now have forgotten strategy please help and be specific. The vector is now linear having been digested with EcoRI and MfeI, then treated with CIAP, to prevent...

23 May 2020 2,377 1 View

Can you reuse PDMS for cell culture?

We have cultured cells on PDMS functionalized with a Matrigel/Collagen mixture, and are wondering if new cells can be directly reseeded after enzymatically removing the original cells. We aren't...

28 November 2018 2,124 3 View

Can I set up a plate for qPCR using Power SYBR Green PCR Master Mix #4367659 and run it a day later?

Catalog number: 4367659 Power SYBR Green PCR Master Mix. This is NOT POWER UP that I am asking about. Can I set up a plate Monday the 28th in the afternoon and run it Tuesday the 28th in...

24 May 2018 8,588 5 View

Does anyone have a tried and true protocol for transfecting RAW cells with plasmid DNA for protein expression?

02 June 2016 7,457 0 View

Does anyone have a complete protocol for generating total cell lysate for proteomic analysis from adherent tissue cultured cells?

04 May 2016 3,844 2 View

How can location be used to tailor news to individual consumers in a way that goes beyond such "lackluster" units of relevance as zip code?

22 September 2015 8,867 3 View

How can I image live cells in 3D systems and subsequently maintain them in culture?

I am growing bladder cancer cells in 3D models in vitro and I wish to visualize them using some form of real-time imaging which will still allow me to grow them and use them for protein/RNA...

11 May 2014 853 7 View

Are there any recent studies on actual carbon emissions within the tourism sector with a breakdown per accommodation type?

The specific data I am finding is quite old and often concerns hotels only and I'm interested in finding out what current emissions are like, particularly in but not limited to Europe.

30 April 2014 1,639 2 View

Solution for patching astrocytes in culture?

I am trying to do patch clamp (whole cell) electrophysiology on astrocytes in culture (namely, primary human fetal astrocytes). It seems for astrocytes, an external solution of pH 7.2 is often...

14 February 2014 8,721 3 View

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

Which is suitable for use with Python? MySQL or SQL Server?

Which is suitable for use with Python? MySQL or SQL Server? What is your suggestion?

01 March 2021 3,422 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 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

Isolates of Hymenoscyphus fraxineus?

Dear Dr. Cai, my name is Simone Prospero and I work in the team of Phytopathology at the Swiss Federal Institute for Forest Snow and Landscape Research (WSL;...

01 March 2021 1,133 1 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 View

Did anyone ever try using pLVX-TetOne Plasmid with a copGFP as a selection marker instead of Puromycin?

When using a lentiviral vector for inducible expression of genes using the Tet-On system. In the literature, I saw a widely used of pLVX-TetOne-Puro Plasmid, is there a reason why people prefer to...

25 February 2021 1,011 3 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View