Can anyone suggest a recipe to make an Electroporation buffer to transfect CH12F3 cells ?

More Lakshani Perera's questions See All

Why there are multiple spikes around 50Hz (Power Interference) in frequency domain (fft)?

I am working on a project related to sEMG classification. In order to process the signal, I am trying to filter the signal, especially power interference. The signal is acquired from Delsys...

07 December 2020 9,745 10 View

What is the best way to plot Ct values for a gene expression in different Samples ?

Hi I want to plot a graph for Ct values from qPCR assay that resulted from a gene expression in different samples.

29 October 2020 1,384 3 View

How to filter resonance frequencies in Matlab?

Following Image presents the frequency domain of sEMG signal which was sampled at 4000Hz. However, unexpected noises were captured during the experiment. I believe that these are some sort of...

29 August 2020 7,421 3 View

What changers can occur if Arsenobetaine (Organoarsenic) digested with an acid (Nitric acid) ?

Arsenobetaine is the non toxin organic form of Arsenic commonly found in fish.If muscle of fish samples subject to a acid digestion, will non toxic nature be changed

30 May 2020 9,109 4 View

Any explanation about the effect of the pH value on SARS-CoV-2 Virus activity ?

Some studies reveal that SARS-CoV-2 virus activity can be mitigated at the alkaline pH medium (above pH= 7.5). Any explanation about the mechanism or the Chemistry behind this phenomenon.

28 May 2020 9,352 3 View

How to input a set of data from an Excel sheet into MATLAB Simulink?

I've got Solar irradiance data and Temperature data for a month in 1 hour time intervals in a Excel sheet. I've modelled a Simulink block set to get the Output of a PV array. Unfortunately, I'm...

27 May 2020 3,072 5 View

What is the best technique / extraction method for extract sesquiterpene coumarins from oligo-gum resins of Ferula asafoetida ?

Phyto-chemicals such as Conferone, Samarcadin, Bdrakemin Farnesiferol A, Fernesiferol C and Galbanic acid can be isolated from Ferula asafoetida. suggest proper extraction methods and solvent to...

22 May 2020 6,350 6 View

Can the sign of a coefficient change from + to - when treat for endogeneity?

Hi, I'm running a model to see the growth effects of the exchange rate. The coefficient that I got for the real effective exchange rate is positive significant under the fixed effect model and...

19 November 2019 6,677 1 View

How do I amplify a 7.4 KB fragment in mouse genomic DNA ?

I am struggling to amplify an 7.4 kB fragment which I need to confirm my CRISPR KO. I have tried Pfu and Phusion polymerase, used both HF and GC buffers, tried 50 - 70 C temp gradient, DMSO...

27 March 2019 9,699 3 View

How to design best primers to amplify a region in mice genomic DNA?

What is the database I should select under the Primer Pair specificity parameters in NCBI primer design tool?

14 March 2019 7,696 2 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

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

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

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 View

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

What is a good reagent for lentiviral plasmid transfection in HeLa cells?

I have been using Lipofectamine 2000 but was hoping there may be a better protocol or one for something like Lipofectamine PLUS reagent.

24 February 2021 1,633 4 View

Why the cells die after stable transfection?

I tried stable transfection in the U87 cell line with the sgRNA-Cas9 All-in-One lentivector. Cells were cultured in a 6-well plate, 4ug plasmid was transfected after 24hs when the cell confluence...

22 February 2021 2,864 4 View

Is there a method for evaluating nanoparticle adhesion to epithelial tissue in vitro?

I want to measure how well nanoparticles can transfect cells based on NP adhesion properties. In the literature I find people do it in animal studies, and epithelial adhesion helps in transfection...

21 February 2021 7,791 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

Saleh Alkarim

Hello

Kindly look at in these links

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2975437/

http://www.jimmunol.org/content/jimmunol/early/2013/07/10/jimmunol.1300922.full.pdf

Good Luck

Christoph Metzner

There's a nice collection of protocols available at http://biorad-ads.com/transfection_protocols/

Best,

Lakshani Perera

Thank you all