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

Calculate pH after addition of base?

04 February 2021 2,232 4 View

Western blot with very acidic proteins, any suggestions?

I am trying to do a Western Blot to detect acidic proteins (~30kDa) that have a PI of ~3.5, due to the presence of Asp and Glu repeats. The proteins are about 80% pure. After I ran them on...

24 January 2021 7,226 3 View

How does one create IFORM contours?

Hello: Can anyone assist with plotting IFORM contours. I have been trying to plot contours using Hs and Tp as variables. The marginal distribution is a GPD-Kernel density and the conditional...

12 December 2020 4,355 3 View

How can I transfer heat between groups in Gromacs?

Is it possible to transfer heat from one group to another using simulated aneeling technique in the Gromacs software? I would like to transfer heat from a gold nanoparticle (group 1 from 310 to...

20 November 2020 9,062 3 View

How to analyze sodium lauryl sulfate using HPLC-RID?

Hi, I am analyzing the content of anionic surfactant: sodium lauryl sulfate (SLS) in commercial detergents using HPLC-RID/refractive index detector. I used column Agilent Zorbax Eclipse XDB C-18...

15 November 2020 1,405 2 View

Seeds of wild Brassica rapa?

Hi, We are a team working on wild and cultivated plants and our stock miss of seeds of Brassica rapa from wild populations in Europe. We are based in Finland, we wonder if any laboratories in...

25 October 2020 857 3 View

Which proteins are continuously expressed (at stable levels) in mammalian monocytes and thus, suitable controls for Protein expression studies?

Hello! Currently I am testing whether an externally added agent affects the expression of only one particular protein or whether it generally inhibits protein synthesis due to its toxicity. For...

14 September 2020 6,359 3 View

Suggestions for an easy experiment on an inverted fluorescence microscope (students lab course)?

09 September 2020 290 3 View

Would ER NiCrMo-3 be the best option for weld overlay of carbon and low alloy steels?

Alloy 625 is the most popular alloy for weld overlay. Without considering the price factor, would NiCrMo-3 be the best technical option for weld overlay of API 5L and low alloy steels, in general,...

24 August 2020 2,478 4 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

What is the state of the drug inside solid lipid nanoparticles (SLNs)?

I am a little confuse with the mechanism of solid lipid nanoparticles (SLNs). The main point of encapsulating drugs into SLNs is because some drugs have low solubility in water. Therefore, SLNs...

01 March 2021 3,703 3 View

MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 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

Can MicroRNA cross the blood brain barrier?

In diseased conditions, many genes and gene products are visible in the bloodstream. So my question is whether we can find traces of microRNA which is involved in disease and disorder in blood serum?

24 February 2021 7,064 3 View

Are exogenous molecules transferred between human cells during doubling ?

Hello RG community, I wonderer whether exogenous molecules (e.g. reporter plasmids, miRNAs or other artificial molecules) are transferred between cultured human cells during their doubling ? So,...

21 February 2021 6,633 3 View

How to obtain free lipids starting from concentration over the CMC?

I need to work with free lipids at increasing concentration (50%, 60% of the CMC ect.). I'm now working with POPC and POPG. To reach these concentrations I have to start from concentration over...

17 February 2021 1,676 3 View

Has anyone encountered any problems with abm good microRNA primers for quantification of mature miRNAs?

I've been trying to quantitate mature microRNAs in 293T cells with and without 'X'. 2ug of total RNA (isolated by Trizol method) is converted to cDNA using All-in one microRNA cDNA synthesis kit...

17 February 2021 8,986 2 View

Solid Phase Extraction of drug product containing nanoparticles ?

Hi Everyone, I am trying to develop an SPE method for the vaccine which contains Matrix-M (saponin Quillaja Saponaria ) which is in nano-particle form and size is around 40nm. These nanoparticles...

16 February 2021 1,389 2 View

What is the best way to fix plasma derived miRNA for subsequent cDNA synthesis?

I need to quantify blood plasma derived miRNA through q-RT-PCR. For this I need to fix RNA content in my cDNA reaction. What is the best way to do this? Currently, I take readings of my plasma...

03 February 2021 9,330 3 View