Why are functional gene results (nosZ, amoA) compared to 16S gene?

I am working on the effect of pH on functional genes in the nitrogen cycle (amoA, nosZ..). Some of the papers i have read present results based on the ratio between functional gene (like nosZ) and...

27 December 2020 1,625 6 View

Why would gel electrophoresis need to be run for 16 hours at 2-8 deg C? Is there an alternative protocol that takes less time or works at room temp?

I am trying to separate and analyze macromolecules (DNA) using horizontal gel electrophoresis. Protocol involves 1X TAE buffer, 10X denaturing buffer, 1X running buffer, 1% agarose gel, and 2X...

18 November 2020 667 2 View

Any tips and tricks to prevent bubbles from being trapped in polymer film during membrane formation via immersion precipitation?

05 July 2017 6,398 6 View

Does anyone know a low molecular weight agarose gel?

I need a trade name for any low molecular weight agarose gel that can dissolve in water. My cDNA is giving me my band of interest (which is faint) and a smear on gel and I want to cut out my band...

23 April 2014 8,122 2 View

Why does the level of phenobarbital spiked in blood appear to be lower than the actual spiked amount when being screened by ELISA?

5ppb of phenobarbital have been spiked into blank blood. This have been replicated by 10 times. However, ELISA results show that all 10 of the replicates contains LESS than 5ppb (average of...

14 July 2013 963 1 View

Can anyone help me with a home made OGD?

I'm hoping to set up some oxygen glucose deprivation experiments using cultured cells but only have a CO2/air incubator. I was wondering whether there was any way of home rigging a small nitrogen...

06 June 2013 2,650 2 View

Will the formation of semicarbazone from an unknown (suspected to be a ketone) be long, or has it been falsely analysed?

In an undergraduate organic experiment, we are required to identify the functional group present via solubility tests (with H2O, HCl, NaOH and NaHCO3) and Classification Tests (Brady's, Tollen's,...

10 March 2013 3,007 6 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

Unknown debris sheets observed in cell culture flasks?

Hi, We culture neural progenitor cells in T175 culture flasks coated with PLO-Laminin. We have successfully grown these cells with our protocol for months. Recently, we observed large sheets...

25 February 2021 3,443 4 View