Why in standard curve qPCR the most concentrate cDNA sample doesn't work and serial dilutions are correct?

Hi! I did a standard curve of my gene of interest. For this, I started with 100ng of cDNA and did serial dilutions (1:10) of this sample 3 steps. The Ct of this samples are: 100ng --> 32,62...

05 November 2019 3,126 3 View

Problems with transfecting fibroblasts?

Hello I am writing because I am having serious problems transfecting foreskin human fibroblasts with several plasmids at a time. At first we started using the transfection reagent Trans-IT-2020...

01 April 2019 1,276 4 View

How to improve efficiency and cell survival in transfection of cortical neurons by CaPhosphate precipitation?

I followed the following protocol (http://www.clontech.com/US/Products/Transfection_and_Cell_Culture/Plasmid_Transfection/ibcGetAttachment.jsp?cItemId=15785&fileId=5875973) using neurobasal...

01 April 2018 2,659 1 View

Have you ever done an F-actin staining with Phalloidin in cells previously fixed (4% PFA) and kept at 4C?

I have a time-course and I would like to start the staining protocol with all the samples at a time

07 February 2018 4,937 3 View

Hi, Does anybody know about how to collect fractions of proteins using FPLC AKTA SYSTEM ?

I mean, to different times of elution and specific pikes of the chromatogram. Thank you very much!

30 November 2016 430 2 View

Does anybody know why in the last step of MSP1D1 protein purification this protein precipitates?

05 October 2014 8,744 5 View

How can I stain for oenocytes in L3 larvae (Drosophila)?

I'm trying to establish a baseline of how fed and starved oenocytes look like in L3 larvae on my wild type strains. But I'm having a hard time finding them. This is how I'm doing the experiment:...

10 April 2014 1,987 1 View

Type of immune response using SRBC or NP-CGG (or similar)

Dear all, I have a naive question. When we want to study Type 1 response, we infect mice with influenza for instance, if we want to study Type 2 response we use for instance T. Bruecei. Does...

01 January 1970 220 1 View

What kind of fluid could I use for a pitfall trap that also does not invalidate molecular testing?

Hi, I want to start testing pitfall trap to obtain ants samples, but I need to conduct molecular analysis on those insects. So, what kind of fluid can I use? Ethanol expires too early and I need...

03 March 2021 5,978 5 View

What are the values needed for gabor filter's parameter for cifar10 data?

Hi, I implemented a code to gabor filter cifar10 data but the images after being filtered and stacked are not clear like the original images. I think the problem is in the way I am using the...

03 March 2021 6,317 1 View

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02 March 2021 3,060 3 View

Would fecal samples stored at -20C for over a year impact metagenomic analysis?

Dear All, I have access to some rodent fecal samples that have been stored at -20C instead of -80C. Anyone have experience with the impact of stowage condition on comparative metagenomic analysis...

01 March 2021 2,512 1 View

How to calculate Total Flavonoid?

I analyzed flavonoid total spectrophotometry, the procedure as below: 2 ml supernatan added 5 ml dd.H2O, then added 0,15 ml NaNO2 5%, 5 minutes later added 0,15 ml AlCl3 10%, After 6 minutes,...

01 March 2021 6,722 1 View

What is involved in morphological changes of SEM images of a substance?

In the sem analysis of CaCO3 nanaoparticles, the morphology images showed spherical shaped particles but the proposed morphology for these type of nanoparticles is trigonal. I prepared the...

01 March 2021 1,960 4 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

How to separate water from glycerol solvent in a solvent recovery vacuum distillation column?

I am currently doing my third year design project. I have been assigned to design a glycerol recovery column in vinyl acetate monomer production. The process unit before the recovery column is an...

01 March 2021 9,525 4 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 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