What sentence do I have to put in the calculation entry to be able to make corrections for overlapping bases (BSSE)?

computational chemistry: I want to make the base overlap corrections for the dichloroacetyl chloride dimer. I put the following sentence to run the calculation in Gaussian: frec B3LYP/6-311++G**...

27 February 2020 6,347 3 View

What binding buffer would you recommend for NI-NTA superflow column purification?

I have to start the purification of a protein using a NI-NTA superflow column. My protein has an aproximate pI of 7. I have no experience with IMAC purification, i know i may have to try...

07 January 2020 3,884 4 View

Why do PC12 cells die after NGF addition?

I have been working with PC12 cells for years. Lately, however, I noticed that they start dying after 3 days of differentiation with NGF . My protocol is the following: I plate the cells onto poly...

02 January 2019 4,794 4 View

What are the best conditions for Western Blotting alpha synuclein?

I am working with PC12 cells and would like to know what are the best conditions to perform western blot of alpha synuclein (14 kDa). Especially, regarding: best buffer for protein extraction:...

08 May 2016 6,236 1 View

What is the best endogenous control for differentiating PC12 cells?

I am working with PC12 cells and I take mRNA samples every 48 hours after differentiation with NGF. Which would be the best endogenous control for qPCR given that it needs to be one that does not...

03 May 2016 2,054 1 View

What are duck egg patterns in inverse distance interpolation, and why it occurs?

25 April 2016 604 3 View

What makes Runge Kutta Method more accurate than Euler method for solving ODEs?

Runge Kutta method gives a more stable results that euler method for ODEs, and i know that Runge kutta is quite complex in the iterations, encompassing an analysis of 4 slopes to approximate the...

02 April 2016 5,304 18 View

What is the function of the ozone scrubber in some ozone analyzers that use UV absorption?

I understand the process of UV absorption, but I am not sure about the Ozone scrubber, is it just a matter of comparison, like a blank for the final result of ozone concentration in air?

22 January 2016 3,473 4 View

What is/are the best agent/s and protocol to differentiate SH-SY5Y cells into dopaminergic neurons?

I want to test effects of a toxic drug on SH-SY5Y cells and I read that there are many agents that induce differentiation into different phenotypes but I could not conclude which of them are...

28 April 2015 754 2 View

Can anyone suggest a protocol for EDS (Ethane dimethane sulphonate) preparation?

04 March 2015 8,537 2 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

How to quantitative analize the data of TNBS assay for gelatin?

Estemeed colleagues, I found some issues regarding the quantification of the data for TNBS assay. There are different protocols on how to perform that but it is clear to me the "fil rouge" that...

02 March 2021 2,616 1 View

Sample dilution for quantitative analysis using ELISA?

If the detection range is in ng/ml but the reference range is in ug/ml for a molecule or protein in serum or plasma .how to dilute and what is the initial volume to be taken for quantitative analysis

02 March 2021 7,670 3 View

How to simultaneously perform multiple biochemical assays?

I am a research scholar working on heavy metal stress on plants. I will be doing biochemical characterisation( protein, carbohydrate, proline, antioxidant enzymes and many more assays) at interval...

01 March 2021 6,999 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 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

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

Why is the efficiency of viral mRNA electroporation in PBMC low?

Hello, I have been struggling with the electroporation of PBMC with HIV-I mRNA. Although the quality of the RNA generated is good, and the handling of the cells in meticulous, the frequency of...

27 February 2021 9,233 3 View

What is the best concentration of cDNA to be used for qPCR?

I have synthesised cDNA from 500ng/ul of RNA. The concentration of the cDNA Is about 5000ng/ul. Should I dilute my cDNA or just use it as it is for my qPCR? I usually use 2ul of cDNA for a 20ul...

24 February 2021 8,618 3 View