What would be the criteria for an expression vector designed for S. cerevisiae to also work in S. pombe?

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What is the recommended/optimal time for resting mouse splenocytes after thawing?

This is my first time working with mouse cells. We isolated and froze down in 90% FCS and DMSO. I would like to retrieve them and do immunophenotyping and intracellular cytokine staining

03 January 2021 6,399 1 View

Can CMOS and CCD images sensors be used as microphones?

I was thinking about Msps and Gsps image sensors in different environments. If a pair of identical image sensors - one exposed to air, gases, plasmas, or partial vacuum - and one in a good...

21 December 2020 9,747 2 View

Is there a tool to find the closest species with an assembled genome for any given species?

As more and more genomes are being sequenced the likelihood of having a close relative being sequenced is rising. So lets say I have a non-model species Chela cachius and I want to find the...

25 November 2020 6,267 4 View

Where can I find a longterm global mean EVI/NDVI dataset?

I am looking for a longterm (10+ years) dataset of mean EVI/NDVI or annual maximum EVI/NDVI and similar. Basically, I am trying to avoid downloading tons of images and doing the averaging myself,...

28 September 2020 8,374 2 View

Can we infuse drug along with sedative?

I would like to infuse rats with drugs of our interest. However, we also need to keep animal under sedation by administrating pentobarbitol via jugular vein with the help of syringe pump since we...

15 September 2020 7,501 5 View

What is a suitable solvent for extraction of arsenic?

Hi. I'm doing some work on arsenic speciation. I am following a paper titled, "SPECIATION OF ARSENIC IN A CONTAMINATED SOIL BY SOLVENT EXTRACTION." which calls for the use of chloroform or benzene...

06 September 2020 6,351 4 View

Ideal saline volume for intravenous injection in rats ?

Hello everyone, my interest is to understand the molecular mechanisms involved in anaphylactic shock using Wistar rats. While i inject non-allergic naive rats with iv bolus (1 mg ova in 1 ml),...

27 August 2020 9,444 2 View

Does mysticism have a scientific basis?

Mysticism is often treated as the opposite of science. But is it? Please see Natural Laws and Mysticism in the Cosmic Cycle of Creation

25 July 2020 6,696 80 View

Which molecular dynamic simulation software is more applicable for protein and large-area surface interaction simulation? And how to run it?

Dear experts: I am trying to do the molecular dynamic simulation for protein-surface interaction, the surface (like cellulose or PET) is a film-like or crystalline material with large surface...

20 March 2020 3,960 3 View

Could our fine-tuned universe have been created from a single fine-tuned quantum particle?

Cosmological explanations for our apparently fine-tuned universe are basically divided between a) a vastly huge multiverse of universes with varying fundamental force and mass constants, including...

21 February 2020 4,618 100 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

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

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

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

Did anyone ever try using pLVX-TetOne Plasmid with a copGFP as a selection marker instead of Puromycin?

When using a lentiviral vector for inducible expression of genes using the Tet-On system. In the literature, I saw a widely used of pLVX-TetOne-Puro Plasmid, is there a reason why people prefer to...

25 February 2021 1,011 3 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View

How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

I have a large 16 kb plasmid, which I need to transfect into PC12 cells. Lipofectamin 2000 didn't work and with GFP alone the transfection rate is very low. I also tried the neon invitrogen...

25 February 2021 6,635 5 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View