Plant research: Watering uniformity/consistency. How important is it?

Trying to gather insight from an unresolved discussion with a colleague here, and either the available literature surrounding this topic and/or my keywords in search of are lacking. Context:...

21 February 2021 6,389 3 View

How to get rough estimation of the concentrations of Oxygen and moisture inside the glove box?

Dear colleagues, I was just wondering if you can suggest a good method to get a rough estimation to the concentration of O2 and H2O inside the glove box as unfortunately, our glove box doesn't...

17 February 2021 1,012 3 View

Onset of Photo Oxidation in Polymers?

Hi! I am looking for some specific information in regards to the minimum amount of UV intensity required to begin the photo degradation process in polymers. I understand each polymer is...

15 February 2021 7,703 4 View

Are you looking for a quality online conference on teaching and learning technology?

This large international conference will be held online this year due to COVID. I am submitting a paper to the Technology Enhanced Language Learning track (8). It is about a topic I have never...

15 February 2021 1,975 15 View

How do you calculate a discrimination index for polytomous data?

Hello, I'm currently running a PCA on a polytomous data set (5 point Likert). I want to create a discrimination index to narrow down my items before running the PCA. After looking around, I'm...

13 February 2021 3,517 2 View

Regarding immunology, transmissivity, herd immunity and vaccines in Sars CoV-2?

This is a follow-up on an earlier question regarding mechanistics of vaccines: why current vaccines don't block transmission. In summary, they don't engage mucosal immune cells to produce IgA....

08 February 2021 8,321 3 View

Which tropical country is the most easy for obtaining research permit (for biological research)?

Dear all, High beurocracy hihly complicate field ecolocial and evolutionary research nowadays. Nearly all tropical countries (maybe all?) requires research permit for performitng research and in...

04 February 2021 3,672 6 View

Can I polymerize NIPPAm with PEGmethacrylate or only with PEGacrylate?

I want to perform the synthesis of a hydrogel and I want to know if I can polymerize NIPPAm with PEGmethacrylate or only with PEGacrylate?

02 February 2021 2,106 6 View

What is the maximum storage time for samples used in soil enzyme assays?

01 February 2021 4,616 3 View

ANOVA vs. T-Test discrepancy?

I have a time series analysis, in which I have determined a mean per each phase. When I run an ANOVA Test with repeated measures comparing the phases, I receive a non significant result, despite a...

31 January 2021 2,960 6 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 4 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

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

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 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