Which contaminative agents must be test before use cell lines for tumor engraft in nude mice?

06 July 2017 1,688 4 View

Why can I not confirm my positive colonies (shRNA ligated in pRNAi vector) via PCR but using a RE can confirm the same positive colonies & plasmids?

For confirmation the cloning of my insert (shRNA) in pRNAi vector, PCR did not respond at all, but when I tested the extracted plasmids of the same colonies with restricted enzyme (scaI), I found...

09 October 2013 1,371 3 View

Can anyone advise on how to clone a small 80 bp segment in a big 4800 bp plasmid?

When I digest the ends of the segment & the plasmid with restriction enzymes, they can not ligate together and although some times I have clones when I use colony PCR I do not have any band.

08 September 2013 2,413 7 View

Why I can not find my insert (80 bp shRNA) in plasmid (5000 bp pRNAi) via colony pcr?

I do not detect my insert in my colonies via colony pcr. What is the reason? I wanted to clone a shRNA in pRNAi. I digested my plasmid & pcr product of shRNA with the same RE. Then ligate them...

08 September 2013 5,209 1 View

How long does it take for tetracyclin to induce the RNAi vector and knock down the gene?

I transfected a cell line with pRNAi vector and then added tetracyclin as an inducer of its promoter. How long do I have to wait if I want tetracyclin to induce the promoter of my plasmid and...

01 February 2013 5,644 3 View

Hela cell line growing slowly

My hela line grows very slowly. I changed my source and got new source of hela line from another institute, but when I passage this new source, the new flasks grow slowly again. what can I do...

31 December 2012 967 10 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

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 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

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

How to simultaneously perform multiple biochemical assays?

I am a research scholar working on heavy metal stress on plants. I will be doing biochemical characterisation( protein, carbohydrate, proline, antioxidant enzymes and many more assays) at interval...

01 March 2021 6,999 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 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

Where to find data on trees in tropical agroforestry?

Hi, I'm looking for data (mainly related to management: growth rate, canopy size, soil and climate preferences, etc.) about tropical trees used in tropical agroforestry. Have you ever heard about...

28 February 2021 7,356 8 View