Can I inactivate restriction enzymes after freezing the digestion product?

I performed a digestion that ended up at midnight, but forgot to inactivate the enzymes (HaeIII and RSAI) before freezing the products. I plan to run a gel to differentiate yeast isolates. Can I...

09 December 2020 3,551 3 View

Frozen spheroid section is not staining (fluorescent + H&E), any ideas?

I have cryosectioned a spheroid which was grown in collagen and agarose and then embedded in OCT Tissue Tek solution before freezing in a dry ice ETOH bath of liquid N2. The sections are very...

08 November 2020 3,299 3 View

Anyone with experience on DNA extraction from biopsies for microbiome analysis?

We shall collect biopsies from skin/hoof of animals for bacteriological microbiome analysis. We need suggestions on DNA extraction kit that may be used for this. We prefer to keep the biopsies in...

24 September 2020 6,845 4 View

How can I remove an article I have uploaded with a wrong title?

24 February 2020 6,806 3 View

Has anyone used a Phalloidin stain that permeates the cell membrane of living cells (rat cardiomyocytes)?

In this particular experiment, it is not possible to perform any permeabilization protocol that weakens the sarcolemma or cytoskeleton, so I would have to rely on finding a Phalloidin that can...

09 January 2018 2,642 4 View

Any tips in conducting the sulfide method using the 'Standard methods for the examination of water and wastewater'?

23 January 2017 3,448 3 View

How do you calculate N2O and CO2 flux emissions collected at different time? What equation(s) would you suggest?

This is a soil incubation experiment in situ. Collection occurred throughout a period of time. Please provide ideas of one or more equations that you could suggest to transform my values into...

21 September 2016 6,857 4 View

Are there any other tracking methods aside from satellite tagging that are just as efficient in tracking the long distance movements of whale sharks?

The migratory patterns of Rhincodon typus is very interesting to me and I have read that satellite tagging is the method typically used when tracking their movements. However, I would like to know...

07 July 2016 3,382 2 View

How to deal with mixed methods studies in mixed methods synthesis?

I have been reading about mixed methods synthesis techniques (Sandelowski et al 2006, 2012 and Joanna Briggs Institute 2014) for systematic review. I think a segregated synthesis approach will...

05 June 2015 138 5 View

What is the minimum fractionation method that is needed for whole mammalian cell lysate prior to Orbitrap-MS?

01 February 2015 9,914 5 View

No title

02 March 2021 3,060 3 View

How to code open-ended questions in SPSS?

Hello all, In SPSS I am going to code 2 open-ended questions. I have already read all the answers and I made a list of the most important categories to which I can code the answers. This question...

02 March 2021 1,757 4 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

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

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

How to eliminate Steady state error from Reinforcement learning genetic algorithm code?

The following code (see 1st 2 images attached) is used to produce PID controller values that are designed to control the system (G). The code finds the PID controller values (noted as k) by using...

28 February 2021 6,560 14 View