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

Mammalian cell culture electroporation?

06 July 2017 6,336 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

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

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 View

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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

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

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

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

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28 February 2021 9,936 2 View

Has anyone had luck using cell lines other than HEK 293 (such as COS1) to package lentivirus?

I am looking to package lentivirus in endothelial cells but I am opposed to using 293 cells due to personal ethics regarding the source. I have done some reading and saw that COS1, COS7, & CHO...

25 February 2021 3,030 1 View