How to test mycoplasma contamination without further contamination of lab areas and pipettes?

Hello I have been asked to test mycoplasma contanimation in some cell culture samples by the adjacent lab using PCR. The lab has been getting their cultures tested outside and know they are...

08 September 2019 4,035 2 View

Can anyone help me with some sequencing analysis files?

Our lab outsourced whole exome sequencing to a company, and I received the analyzed files recently. I have always been a wet lab person, and do not understand most of the file types, and how to...

02 March 2018 6,602 5 View

What is the best marker to study Vascularization in Human cortex using RT-PCR?

We are interested in looking at vascularization and angiogenesis in some human brain samples. The ideal way would be to use immunostaining but our lab is not developed for IHC or any...

05 June 2016 3,451 2 View

Has anyone used the Biorad multiscreen apparatus with a 10-well comb/run?

08 September 2015 9,001 1 View

How do you account for cell confluency when performing knock down studies in differentiated vs undifferentiated cells?

I am trying to knock down a gene of interest using siRNA in SH-SY5Y cells. But the amount of siRNA I use is mostly based on the well surface area etc. I use 50pmol/well in a 6-well plate. So is it...

02 March 2015 8,753 4 View

How can we isolate single-cells after transfection to generate stable SH-Sy5Y cell line? Also, do these cells grow well from a single cell?

I will be using CRIPSR genome editing to modify a gene and generate stable cell line. I intend to use a plasmid with puromycin selection marker to initially screen the cells. Does anyone has...

31 December 2014 183 7 View

Any suggestion for a transfection reagent for Sy5Y cells for siRNA transfection?

Hi, We have used FuGENE 6 (Promega) to transfect Sy5Y cells with plasmid DNA, but I just spoke to the promega tech support and they do not recommend FuGENE products or any of their transfection...

10 November 2014 3,722 3 View

When I start a culture of COS7 cells from stock, can we use the cells for transfection after growing them overnight?

10 November 2014 8,541 4 View

To transfect SH-Sy5Y cells, do we need to start a culture from new frozen stock and wait 3-4 passages every time before doing transfection?

Can we not transfect the cells straight from Liquid N2 after culturing them overnight or something like that? Has anyone ever tried this?

09 October 2014 5,257 2 View

What solvent can I used for Paal Knorr reaction between 6-amino hexanoic acid and 2.5 hexanedione?

I am try to make the Paal Knorr reaction between 2.5 hexnedione and 6-amino hexanoic acid, my problem is type of solvent to use, because 6-hexaminohexanoic acid is soluble in water but I am not...

02 March 2021 4,443 2 View

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02 March 2021 3,060 3 View

New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

02 March 2021 3,024 2 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 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