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 use Quinolinic acid for cell culture treatment?

I want to design an experiment where we treat the neuroblastoma with quinolinic acid. Has anyone done this? Or treatment of primary cultures with QA? Where do you buy QA? And how do you suspend...

04 May 2015 4,962 2 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

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

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

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02 March 2021 3,060 3 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 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

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