Anyone knows difference between Primescript and MMLV RT enzymes?

Hi Everybody, Still questions about PCR for circRNA. I do some review for papers and found many of them use MMLV for DNA systhesis. Well in my lab, we usually use Primescript from TAKARA. and I...

04 May 2019 6,808 2 View

Any drugs or chemical can inhibit sox2?

Hi All, I'd like to inhibit sox2 express or down regulated in my current cell culture, but shRNA method seems hard to apply as my cell line is a specifial neuron pregenitor and very hard for...

06 July 2016 6,089 1 View

Colocalization analysis for 3 channels ?

Hi All, I have question about colocalization analysis for 3 channels. I attached an image to make my questions more easily understand. I have 3 images by confocol, from 3 channeal, green, red and...

06 July 2016 9,988 5 View

Self-made electrode, what materials did you guys use?

03 April 2016 8,649 2 View

Subclone PCR, what tag or method can I use to generate long insert piece?

Hi All, My current work needs to insert a large piece DNA into a vector , it's about 5k bp long. We were used invitrogen HF-tag for PCR many years, and it works very good for 1-2k bp insert PCR,...

03 April 2016 6,686 3 View

Did anyone do the electroporation on neurons? or neuron stem cells?

02 March 2016 2,482 4 View

I have RNA integrity gel 2 bands but they are the wrong size.

31 December 2015 8,653 17 View

Why do I get bright GFP-reporter signal but no increase in the mRNA levels?

Hi everyone, I really get a weired problem, I used a plasmid transfection cells, the plasmid has a GFP reporter linked with a protein (lets call it A protein) which I was interested, after...

10 November 2015 3,377 5 View

What if I add another cDNA sequence after the stop code, will it express protein?

Hi All, I was thinking about in translation part, stop code will stop translation, and ATG will start translation, so what if I make a plasmid, it has promoter, then a cDNA start with ATG and add...

03 April 2015 4,878 6 View

Can I put 2 promoters together and hope both of them will function?

Hi All, As my problem, I am working with ES cell lines and want to induce them into neurons. in this process we want to observe it as GFP+ cells. but seems the promoter (i.e ef1a) express high in...

03 April 2015 1,093 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 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

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

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

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 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