Has anyone used PolyJet Transfection reagent with Stealth RNAi?

More Mandy Kain's questions See All

Hindquarter infections in BALB/c mice

A few of my male Rag 2 -/- BALB/c mice have gotten infections - bilobed masses on their hindquarters. Many of the mice do get rectal prolapses, but these infections have been occurring in mice...

31 December 2012 4,452 3 View

Stimulating Treg production?

I would like to culture splenocytes in order to expand their Treg population using TGF-B, CD3 antibodies, and CD28. All the protocols I've found are for pre-sorted T cells using an isolation kit...

08 September 2012 4,064 8 View

Tissue-specific tumor markers

One of our mouse strains has developed large tumors in their necks that may come from either the mammary glands, the salivary glands, or the thyroid. Do you know of any markers specific for these...

05 June 2012 9,654 13 View

Transfection/Infection background sources?

I'm trying to create a knockdown line using either viral or non-viral methods (depending on availability), but I've never done this before and have only used established cell lines. Does anyone...

04 May 2012 10,011 4 View

Click-it Protein Synthesis Assay from Invitrogen

I'm using the Click-it Cell Reaction Buffer Kit with AHA azide and AlexaFluor 647 alkyne to measure protein synthesis in HeLa cells and I'm having trouble reducing the background. The protocol...

31 December 2011 9,090 0 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 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

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 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

What is the best concentration of cDNA to be used for qPCR?

I have synthesised cDNA from 500ng/ul of RNA. The concentration of the cDNA Is about 5000ng/ul. Should I dilute my cDNA or just use it as it is for my qPCR? I usually use 2ul of cDNA for a 20ul...

24 February 2021 8,618 3 View

John Williams Popular answer

Technically speaking RNAi can work with plasmids; as long as they encode shRNA, it will be produced within the cell and enter the RNAi pathway. Regardless, siRNA transfection generally has higher efficiencies (see examples here: https://altogen.com/products-index/) and is easier to execute. Library screening to determine siRNA specificity is a common way to find what works best for silencing your target gene.

John Williams