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

Is there a method for evaluating nanoparticle adhesion to epithelial tissue in vitro?

I want to measure how well nanoparticles can transfect cells based on NP adhesion properties. In the literature I find people do it in animal studies, and epithelial adhesion helps in transfection...

21 February 2021 7,791 3 View

Can qPCR used to quantify cells over other methods like ICC & flow cytometry?

I am currently working to generate a certain type of neurons. However not all the cells in the culture will be expressing the specific markers. I would like to check how many cells are expressing...

03 October 2020 4,753 2 View

Is Heat Inactivation of FBS always required before adding to medium?

So I have done once Heat Inactivation of FBS which is from an entirely new bottle, and then I prepared few aliquots with 5mL each. These few aliquots of FBS are most probably to be used within 2-3...

01 July 2020 3,822 3 View

Hek293 T cells contamination from CO2 incubator or media/serum?

Dear all, Hello! I have had issues with Hek293T cells maintenance for quite awhile whereby my Hek cells die after passages 6 or 7. I've recently started trouble shooting to find out the source of...

09 June 2019 6,487 7 View

Can Solidworks Cosmos/ ANSYS Fluent be set to simulate (steady-state thermal analysis) only for few hours of real time?

During experiment, a pyrolysis reactor was constantly heated with 825 Celsius of heat for 2 hours with other parameters remained constant. How can I set in Solidworks Cosmos/ ANSYS Fluent to...

05 November 2017 7,283 2 View

How to check for gender differences for multiple regression in SPSS?

I am running a multiple regression analysis for 2 predictors (parental conditional positive regard, parental conditional negative regard) and 1 DV (resilience). All are continuous. I wish to...

02 September 2017 2,171 10 View

Sectional analysis on UHPC beams

Hi, I am attempting to carry out sectional analysis to determine moment capacity of UHPC beams. I carried out 3-points bending tests to determine the flexural strength of the material. However I'm...

04 July 2017 3,459 3 View

Can anyone recommend me any human cell line for transfecting FOXO3 gene?

Hi, I want to overexpress a gene named FOXO3 in a transfectable cell line. Can anyone recommend me any human cell line to do that?

09 June 2016 4,842 1 View

How can I predict risk of road accidents (mathematical models) through the combination of driver behaviour questionnaire (DBQ) & Instrumented vehicle?

27 April 2015 1,145 7 View

Does the pressure(-600 mmHg) affects pH?

When I conducted the degassing experiment using decompression under anaerobic condition(and I used ferrous based chelating agent), the oxidized ferric were observerd. So, could you tell me the...

03 March 2021 4,892 5 View

What is the production process of CaFe ferroalloys?

I looking for ferro calcium production process

03 March 2021 1,755 4 View

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

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

Which database would give me this list of chemical components?

Dear colleagues: I would like to know if anyone knows of a database that gives me the parameters pH, K, K, Mg, P, C, N, Zn, Cu, Ni and Cd of common foods. I find a lot of information of proteins,...

26 February 2021 5,525 3 View

Unknown debris sheets observed in cell culture flasks?

Hi, We culture neural progenitor cells in T175 culture flasks coated with PLO-Laminin. We have successfully grown these cells with our protocol for months. Recently, we observed large sheets...

25 February 2021 3,443 4 View

What is a good reagent for lentiviral plasmid transfection in HeLa cells?

I have been using Lipofectamine 2000 but was hoping there may be a better protocol or one for something like Lipofectamine PLUS reagent.

24 February 2021 1,633 4 View

Why is NH4OH added in heavy metals limit test?

The limit test I'm referring to is USP method II. In this procedure you need to dropwise add 6N of NH4OH to the sample until it the pH is alkali. After that you need to dilute and add HOAc until...

23 February 2021 2,049 3 View

Is there an ideal incubation time of RGO-modified SPCE before subsequent use, i.e., electrochemical characterization or further modification?

I notice that upon washing and air-drying of my RGO/SPCE surface and characterizing again via DPV, the current response towards ferricyanide probe significantly decreases. My worry is that the...

22 February 2021 5,324 3 View

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22 February 2021 9,932 11 View