How can I do RNAseq?

I have murine B cells. I'd like to perform RNA-seq upon treatment. How much starting RNA is required to do it?Could someone shares a protocol? thanks in advance Simone

09 October 2015 6,971 0 View

What program do I have to use for Daudi transfection using AMAXA?

Hi guys, could you share with me your protocol to transfect DAUDI cells by AMAXA? I have to do siRNA transfection. I have the nucleofector solution V thanks

11 December 2014 4,260 1 View

Can someone suggest a protocol for SiRNA and suspension cells?

Hi guys, I'm going to perform a KD of my protein of interest. The problem is that I'm working with suspension cells. Could someone suggest a protocol avoiding electroporetion? thanks

09 October 2014 5,104 6 View

How is it possible to eliminate polymers?

Recently I observed polymers in my samples. Do you have some suggestions to avoid their presence?

04 May 2014 8,761 7 View

Can anyone give me suggestions on doing nuclear/citoplasm fractionation?

Unfortunately when I performed N/C fractionation I assessed protein precipitation. Usually I used a hypotonic buffer to separate C from the N. On the nuclei I applied a strong saline buffer. The...

03 April 2014 3,542 4 View

Do you know which markers of Euchromatin and Heterochromatin I could use for IF?

I'm interested in understanding if my protein will colocalize with chromatin.

02 March 2014 7,775 4 View

Any suggestions for the fractionation of suspension cells?

I'm having a lot of problems performing nucleus/cyto fractionation in DAUDI cell line. The protein recovery is really low. Do you have any suggestions?

02 March 2014 2,812 3 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

No title

02 March 2021 3,060 3 View

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 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

Could you please recommend some good journal that accept very long revie papers (approximately 6000+ words) in neurosurgery, cancer biology ?

Could you please recommend some good journal that accept very long revie papers (approximately 6000+ words) in neurosurgery, cancer biology ?

01 March 2021 8,087 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