Single E coli colony transformation?

Is there any protocol to make chemically competent E coli from a single colony and transform it? Any buffer recipe? Many thanks

02 March 2018 9,655 0 View

Electrocompetent bacteria storage at -20?

Hello, I'm trying to make electrocompetent bacteria and store them at -20 degrees as I don't have -80 freezers. Is there any protocol? Should I use 50% glycerol? Thanks

06 July 2017 3,030 1 View

Eukaryote Promoter consensus sequences analysis ?

10 November 2016 936 4 View

DNA "Pellet-Paint" coprecipitant or similar composition?

09 October 2016 8,164 9 View

Tumoral cell line intradermical nude mice inoculation?

Hello, I need to generate tumors in nude mice with the cell lines SK MEL 28 (melanoma) and MDA MB 231 (breast cancer), both human tumoral cell lines. How many cells do i have to inject...

05 June 2016 3,257 4 View

PEI transfection of SK-MEL 28?

02 March 2016 5,205 4 View

Has anybody got any protocol to make competent lyophilized E coli?

Hello, Has anybody got any protocol to make competent lyophilized E coli?

10 November 2015 1,950 0 View

Add ssDNA to a Gold Nanoparticle?

09 October 2015 3,167 1 View

Protocols to liofilize DNA?

05 June 2015 8,525 3 View

What is an equivalent alternative reagents composition like the Zymo Mix&Go?

Searching the web i found that Zymo Research makes a kit for making competent cells and transforming them in aprox 20 seconds without incubating the mix on ice for 30 minutes, without heat-shock...

04 May 2015 3,463 4 View

No title

02 March 2021 3,060 3 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 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

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 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

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

What could be some reasons why unstimulated monocytes are getting activated?

Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.

28 February 2021 7,883 3 View