optimize the condition?

More Dyaningtyas Dewi Pamungkas Putri's questions See All

How to solve problem in over expression research?

Hi.. Recently I do over expression some gene using plasmid target and transfection in hek 293 cells. My plasmids are flag pcDNA3 and pCMV-myc plasmid. When I did that experiment then I analyze...

01 February 2017 4,295 2 View

How to measure phosphoinositide in cell line?

I'm working on the cell lines and want to investigate level of phosphoinositide PtdIns, PtdIns (3), PtdIns (5), PtdIns (3,5), PtdIns (3,4,5). Some paper said using the radiolabbeling but I would...

10 November 2016 2,644 1 View

How poly IC in transfection?

Dear all, how microliter of DNA 1ug/ml I should apply in volume of dilution medium to get the final concentration is 1ug/ml in 96 well, 24 well and 6 well Thank you for your help

04 May 2016 9,460 2 View

Does anyone have experience in neutrophil movement?

i would like to investigate the movement of neutrophil in 96 well plate and i need to coat the well. does anyone have experience about the best coat for neutrophil movement? BSA? HSA? FBS?...

01 February 2016 8,422 4 View

Why do I have problems in transfection?

i check the condition of the cells. But I get the worst condition the cells. Only a few cells that attach on TCD. so, may you help me to solve electroporation problem? thanks

08 September 2015 3,761 6 View

What is the critical step when stimulation?

in my case, may I say that the production of IL10 of the supernatant isn't enough? Is it necessary to increase the cell number? Thanks for helping regards, Dewi

04 May 2015 9,551 1 View

Does anyone have some experience in PCR with gene length about 4,000bp?

I try to PCR my gene target, that length about 4,000 bp. First time PCR, I try with cDNA 5ng, each concentration primer 0.1mikroMolar, dNTP 0.4mM, KOD FX 1.0U. and I got the smear band, I try to...

01 February 2015 8,452 22 View

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

Why activated CAR-Jurkat cell could not kill targets?

Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...

06 August 2024 641 2 View

Are there instances where molecules with larger molecular weights exhibit greater mobility than those with smaller molecular weights?

Hi, I know that low molecular weight (MW) molecules generally tend to have higher mobility, while high molecular weight molecules tend to have lower mobility. However, in my experimental...

06 August 2024 1,495 2 View

How much total RNA concentration to be extracted from sorted plasma cells from bone marrow of C57BL/6 mice for RT-PCR ?

i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...

05 August 2024 8,835 1 View

Weak DAPI staining after immunohistochemistry - how to improve?

After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...

05 August 2024 9,637 2 View

The Curse of Evolution and Complexity?

Brain and body mass together are positively correlated with lifespan (Hofman 1993). The duration of neural development is one of the best predictors of brain size, and conception is the best...

05 August 2024 6,247 3 View

For an in-vitro drug release study, what molecular weight cut-off (MWCO) dialysis bag is required for a 117 kDa protein?

kindly reply me. Thanking you in advance.

05 August 2024 7,727 4 View

How to use Density Functional Theory to calculate carrier mobilities of solid system?

Hello, everyone. I have tried to determine carrier motilities of some materials, by Density Functional Theory, using Quantum ESPRESSO. There are a few methods to do it, like a package called...

04 August 2024 8,894 1 View

Does long-term culture of murine Monocytes result in macrophage differentiation?

Hi, I am isolating monocytes from the bone marrow using the Mouse Monocyte EasySep kit. I want to treat these cells and monitor expression of specific markers over the course of 10 days. I will...

04 August 2024 7,282 2 View

How to start a Molecular Dynamics Simulation?

Is it possible to conduct a molecular dynamics simulation to see the effects of a specific carbohydrate on the structure of lipids (e.g., micelle structure)? I am a beginner in this field and plan...

03 August 2024 3,371 3 View

Sreedhar V

below link may provides useful information.

https://www.neb.com/tools-and-resources/usage-guidelines/electroporation-tips

Hope this helps little

Refik Kanjhan

Dear Dyaningtyas,

I usually do very succcesfull single cell electroporation of dyes and large molecules (mRNA and genes) using microelectrodes. I think it may be useful for you to understand how electroporation works at the electrode-membrane interface of a single cell. How the membrane breaks and re-seals (repairs) following voltage pulses. I recommend you to read my detailed methodological paper listed below.

best wishes, Refik

Article Semi-loose seal Neurobiotin electroporation for combined str...