What's the longest insert you put into a plasmid via quickchange-like methods?

We would like to put a 12-20 aminoacid linker into a recombinant protein expressed in E coli. We've had good experience with Quickchange-like protocols; but never tried with 36-60 nucleotide...

02 March 2019 3,185 4 View

Is Lithium Lauryl Sulfate OK to run K+-containing samples on SDS-PAGE?

Hi all, Can one use Li Lauril Sulfate (LDS) in loading buffer to run protein samples containing high concentrations of KCl, on SDS-PAGE? Does one need to replace SDS in the running buffer and in...

01 February 2019 1,614 6 View

Is there a chemically defined, FBS-free media formulation for HCT-116 and U2OS cells (adherent human cells)?

FBS-free medium is what I am looking for. If defined growth factors are required, I can express them (if they are available in recombinant form). Artificial meat companies seem to have figured it...

06 July 2018 7,101 4 View

Suggest a method for harvesting of adherent cells from hollow fiber modules? Does scraping work? Would using hydrophobic fibers help?

We are looking for a cost-effective way to grow adherent cells (HCT116, US2OS), in amounts of 1*10^10, for subsequent isolation of nuclei and preparation nuclear extracts per Dignam 1980....

05 June 2018 7,168 5 View

Has anyone tried using standard hollow fiber PES hemo-dialysis cartridges to culture adherent mammalian cells?

The surface area (1-2 sq meters) and the basic specs (excluded volume, PES memberane chemistry, pressure drop, non-fowling, sterile, QC, Mw rejection) of hollow fiber dialysis filters appears...

05 June 2018 9,437 0 View

Can 5+M guanidine-HCl denature dsDNA PCR products upon prolonged exposure (>16 hours, room temp, this is for crush and soak of acrlamide gels)?

I am wondering if prolonged incubations in 5M GuHCl when crush-and soaking acrylamide gel slabs can denature DNA. I normally use water or the Maniatis buffer, but having 5M GuHCl around would help...

02 March 2018 7,428 3 View

Does anyone Happen to know what BioRad's iProof PCR enzyme is (beyond what's in the catalog description)?

09 October 2016 1,608 2 View

Does anyone have experience with the mini8 PCR cycler?

09 October 2016 3,488 0 View

How to control pH and composition of PEG solutions?

Hi all, I am trying to concentrate a DNA sample using dialysis against 50% PEG, through a 10K cut-off membrane. I made a 50% solution of PEG20K supplemented with 10mM Tris (target pH=7.8), and...

08 September 2016 3,873 10 View

An affordable and reliable thermal cycler?

02 March 2016 3,750 5 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

Why can't make aluminum nitride film successfully by use sputter?

We use reactive sputtering process to make aluminum nitride film, and want to make c-axis aluminum nitride film on a silicon substrate, but the crystal orientation cannot be seen with...

03 March 2021 7,975 3 View

Is there any test to determine the cardiovascular risk of patients with obesity? from the perspective of obesity?

We found tests to determine cardiovascular risk taking into account lipid profile, blood pressure, age, history of diabetes, other laboratories. But when weight is involved, there is a paradox. I...

02 March 2021 6,036 1 View

Why are the velocities so high in my Boussinesq approximation Fluent simulation?

Hello, I have a mechanism where air rises due to convective flows. In this simulation, nothing is driving the air except this. I understand that the Boussinesq approximation only works for small...

02 March 2021 4,910 3 View

Why almost every element would be metallized in sufficiently high pressure and low temperature?

(This statement is from wikipedia, BTW). What is so special about metallic bond that is limiting high-P low-T state of a matter (as long as individual atoms exist, not white dwarf of neutron star...

02 March 2021 3,309 2 View

Sutter electrode holder leaking after applying positive pressure. How to fix?

I am using a Sutter electrode holder for my patch clamp experiments, but I'm having trouble keeping positive pressure inside my electrode due to a leak out of the back of the holder. I have added...

02 March 2021 5,829 1 View

What SPSS analysis should I run with one IV (with two levels) and two DVs?

Hi, I am trying to figure out which test would be most appropriate to run for the following question: Is there a difference in task blood pressure between smokers and non-smokers? There are two...

01 March 2021 8,235 2 View

How much pressure we can reduce at the outlet to avoid Reverse Flow?

I am simulating Heat Exchanger. I have tried and tested all the methods to resolve this issue from the internet like refining mesh, improving skewness and orthogonal quality ,extending outlet to...

01 March 2021 6,985 3 View

For gelatin coating for culturing HUVEC.Should the gelatin be made in sterile water or PBS? Do I need to autoclave the gelatin soln after making it ?

I want to use 0.1% gelatin. Should that be made in PBS or sterile water? Should I autoclave the gelatin soln after making it in water/PBS?

28 February 2021 274 2 View

The reason that the culture of white fungi Contamination with green mold(Trichoderma), the culture autoclave at121 for 15 min. What is it?

Is the period to autoclave not enough? The inoculation in a hood and flame and UV and alcohol.

27 February 2021 9,356 3 View