How optimize the Size Exclusion Chromatography?

Hello! I'll do a size exclusion chromatography, but I only have an open column, and I'll perform the peptide extraction from yeast, using buffer lysis (sodium phosphate 50 mM/NaCl 30 mM/DNAse and...

01 March 2021 2,215 2 View

What thickness need for ITO to use as conductive layer?

I want to do RF sputtering of ITO on glass substrate using magnetron sputtering. what thickness do i need to make it good conductive layer to use it for PSC. Moreover, whats about thickness of...

01 March 2021 395 3 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

Special issue on PLS-SEM in Mathematics. Are you interested?

21 February 2021 5,601 3 View

How to calculate the TCID50 when the highest CPE is 50% or less?

I am doing TCID50 to measure progeny virus and some treatments protected the cells in a way that the highest CPE observed is 50% or less. How should I do the titre calculation in that case? Thank you!

16 February 2021 8,768 1 View

Are there any differences between setting automatic or manual number of measurements in DLS?

When doing measurements in DLS (either size or zeta potential), I have the option to input an automatic number of measurements or to set it manually. My question is: is there a drawback in...

08 February 2021 9,423 6 View

Which drug is better to modify blood pressure in awake monkey, if I provide it in the food or drink reward?

I need to modify the blood pressure in a monkey while is awake in the recording chamber, and I am planing on provide a drug for that in the food or water used for reward... any advise? Thank you

08 February 2021 9,900 1 View

Why stop the reaction in ice for the fluorescent probe FDA (fluorescein diacetate)?

Hi, In literature I noticed that who stains haemocytes with FDA for 30 min then stop the reaction in ice while this does not apply for 15 min incubation time (or maybe authors did not write this...

08 February 2021 3,366 3 View

How to make Fast Green FCF from powder?

Hi everyone We need Fast Green to electroporate plasmid into mice embryos, therefore we simply need it to see some color in the liquid we are electroporating so that electroporation is more...

07 February 2021 1,671 2 View

Why do high doses of humic substances inhibit the growth of corn roots? Is there a toxicity effect?

Length, volume, surface area and dry mass of corn roots treated with humic substances increased up to 13 liters per hectare and decreased with higher doses.

02 February 2021 2,190 4 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

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