PCR using oligo dT?

Is it possible to perform a PCR, using as the reverse primer a oligo dT, the same used in the reverse trancription? I am trying to amplify a mRNA, but with no success

21 June 2018 3,707 3 View

Denaturing RNA gel

Does anyone has a denaturing RNA gel protocol, by just adding guanidine to the sample, and running in a regular agarose gel?

19 July 2017 8,078 3 View

How do you do the normalisation of X protein to study the effect of a protein Y mutation on X protein level by western blot?

I need to study the effect of the protein Y mutation on the levels of a protein X. So I over-expressed the protein X under 5 conditions: (1) not over-expression of protein Y (2) with...

07 June 2017 5,252 4 View

Is it possible to load 120 ug of proteins in a protein gel?

I need to load 120 ug of proteins in a protein gel. Do I need some precaution for the protein migration and western-blot?

11 July 2016 624 4 View

Convolutional Neural Network: How to increase input resolution in a CNN?

24 February 2016 5,026 3 View

High variability in western blot, why?

I am trying to detect endogenous Aurora B in the whole cell extract but I have high variability during the detection by LiCOR scanner. I use 2 different primary antibodies from BD technology and...

10 December 2015 4,007 6 View

2-Amino-2-norbornanecarboxylic acid (BCH) is soluble in PBS or MEM medium?

I should use the BCH for inhibition experiment for LAT1. I saw that BCH is soluble in 1M NH4OH but I cannot use this solution with cells.

07 April 2014 6,162 3 View

How can I digest nanorods to measure gold with ICP?

I bought nanorods from Nanopartz and I want to measure the gold. I tried to digest the same amount of nanorods with 4ml of aqua regia for 6h at 100 celsius but every time I did, the ICP resulted...

10 February 2014 4,343 2 View

Does anyone know the Bio imaging navigator from Olympus (FSX100)?

My lab is interested in this all-in-one microscope system and I'd like to know if someone has already used it?

07 August 2013 5,737 1 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 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

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

Help request in CST microwave?

im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...

02 March 2021 2,246 2 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

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 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

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 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

Dapi fluorescence protocol for colon cell lines ?

28 February 2021 2,534 2 View