Co-transfection of miRNA mimics and inhibitors?

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Transfected cells died after trypsinization ?

Hi all, I am facing a problem to trypsinize and combine/move the transfected cells into new well . I was planning to pool the transfected cells into a new well to ensure better cell-cell...

11 December 2019 3,243 5 View

Constitutive expression of eGFP in transfected cells?

Hi, I am trying to culture a stable clones using transfected cells. As the clones grow, I noticed the eGFP signal diminishes over time but the cells are still resistant to antibiotics and do not...

11 December 2019 176 14 View

New in lentivirus transduction work?

Hi all, I am new in lentivirus transduction work and have some problem as listed below: 1) Is CMV promoter considered a strong promoter to express the gene I want in the transgenic organism...

09 October 2019 8,592 4 View

Problem with stable cell line generation?

Hi all, I am facing a problem to generate a stable cell line after successful transfection. I added puromycin to kill the untransfected cells, however, eventually all my transfected cells are...

08 September 2019 9,024 2 View

Will miRNA transfection affect cell morphology?

Hi Scientist, I am transfecting an oncogenic miRNA into a breast cancer cell line and based on your experience in the mammalian cell line, will such transfection affect the cell morphology?

06 July 2019 7,378 8 View

Suggestion for good plasmid extraction kit?

Hi, Can anyone recommend a plasmid extraction kit that could produce good plasmid yield (>300ng/uL). The plasmid I am using now is a high copy number plasmid and the bacteria I am using is...

06 July 2019 6,820 3 View

MiRNA primer sequence?

Dear all, I have a miRNA primer sequence left by someone else and I would like to know about the sequence of this primer. Is there any method I can use to sequence and know the sequence of this...

05 June 2019 5,879 7 View

Use of fluorescence or luciferase approach in checking transfection efficiency?

Hi all, I would like to seek for your opinions on whether fluorescence approach (GFP) or luciferase approach is better in assessing transfection efficiency / success (after miRNA...

03 April 2019 1,383 6 View

Single plasmid that carries both miRNA and reporter gene like GFP/luciferase?

Hi researcher, I wonder any of you have tried to design a single plasmid that carries both miRNA and reporter gene like GFP / luciferase? I am thinking with such plasmid, I will just need to...

03 April 2019 5,144 7 View

Viral or non-viral constructs for transfection?

Hi all, I would like to ask your opinion on whether viral or non-viral construct is better and more superior for transfection involving human breast cancer cell lines, MDA-MB-231? Besides, in...

03 April 2019 6,372 4 View

What element is driving the different slop in the low frequency tail?

My study consists simple corrosion inhibitors in salty brine. Right now I just fit my circuit with randles with CPE. Just wondering how the slop can given more info of the experiment. Or any...

01 March 2021 6,474 3 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...

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MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 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

Can MicroRNA cross the blood brain barrier?

In diseased conditions, many genes and gene products are visible in the bloodstream. So my question is whether we can find traces of microRNA which is involved in disease and disorder in blood serum?

24 February 2021 7,064 3 View

Can I use a protein-devoid condition as my negative control in ELISA?

Hello, I am running a sandwich-type ELISA where I am trying to inhibit a protein-protein interaction using small molecules. For each experiment, I determine my Z' (Z-prime) to determine the...

23 February 2021 7,429 1 View

Alexander Sasha Vlassov

for the same miRNA target? never saw such a paper, everyone is transfecting either mimic or inhibitor- separately, so that you can either increase or decrease specific miRNA levels.

Zx Chong

Yes Dr. Alexander Sasha Vlassov . I was recommended by some people on transfecting both miRNA mimics and inhibitors in the same cell.

You are right. It is hard for me to find papers that reported co-transfection of mimics and inhibitor in the same cell.