How to check miRNA transfection efficiency in case of buffalo endometrial epithelial cells?

More Ankit Pal's questions See All

Why is the sum of the elements in a row in a Q matrix equal to zero?

When explaining substitution models, the substitutions are expressed as Q matrix. Why is the sum of the elements in a row zero?

28 February 2021 3,864 3 View

What is the method of removing RNase contamination from a filter paper surface?

I am working on a biosensing project on paper substrate, however, it is found that the RNase contamination occurs on the filter paper. Might I ask is there any method to clean the paper surface...

12 February 2021 3,926 3 View

What is causing the raw fluorescence to be negative?

Hi everyone, I would really appreciate it if someone can tell me when and why a raw fluorescence starts below zero. In the picture attached below the blue colour curve starts from a negative...

09 February 2021 4,642 3 View

Regarding immunology, transmissivity, herd immunity and vaccines in Sars CoV-2?

This is a follow-up on an earlier question regarding mechanistics of vaccines: why current vaccines don't block transmission. In summary, they don't engage mucosal immune cells to produce IgA....

08 February 2021 8,321 3 View

How can I enter the load increment values to the Abaqus?

In Abaqus I know that there are two methods for incrementation (Automatic and fixed incrementation). I want to control the incrementation by myself. I want to input the values of load increment...

03 February 2021 6,943 3 View

Desalting using DSC- 18 SPE columns?

I am looking forward to remove excess of salt form my metal based porphyrin and trying DSC-18 columns for the same, can someone suggest me how how can one decide the polarity of the solvent to...

01 February 2021 9,076 1 View

Distortion problem in Abaqus explicit?

Dear all. I'm modeling the RC column with cyclic displacement by Abaqus-Explicit. The concrete model used Concrete Plastic Damage model. However I'm facing with a error about the excessive...

01 February 2021 7,726 1 View

I am finding a lot of cellular debris in the culture flask? What are the reason behind it?

I am facing a problem that cellular debris observed in my SHSY5Y cells. I have seeded the cells in DMEM media with 10% FBS in T-25/ 60mm dish (antibiotic used PEN-Strepand gentamycin). I have also...

17 January 2021 3,074 3 View

Can we use two antibodies generated in same species for indirect Sandwich ELISA?

I have generated two antibodies against two peptide fractions of a protein in the rabbit. I want to use these two antibodies (Ab-1 and Ab-2) in indirect sandwich ELISA for the detection of...

11 January 2021 1,754 4 View

What is the reason for discontinued Stage II fatigue crack growth in pearlitic steel?

Conventionally, the process of Stage II fatigue crack growth is governed by Paris law with less microstructural effect. When fatigue crack growth test was carried out for pearlitic steels ( both...

03 January 2021 7,075 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...

28 February 2021 4,949 3 View

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?

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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

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

Saurabh Mandal

if it fits with your study context, you can think of a 3'UTR luciferase reporter assay.

John Hardy Lockhart

One thing I'll point out is that the use of qRT-PCR to assess transfection efficiency is not a direct measure since the efficiency of mRNA knockdown is also highly dependent on the efficiency of miRNA mimic binding.

To directly assess transfection efficiency I would recommend using a fluorescently labeled siRNA control and measuring using flow cytometry.