Is GC content important when designing oligonucleotides for transfection?

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Are primers used in reverse transcriptase PCR also used in real time PCR?

Are primers used in reverse transcriptase PCR also used in real time PCR? Or should I redesign the appropriate primers? If so, what is the difference?

11 December 2019 1,822 4 View

I cannot detect Parp protein after total protein isolation?

I cannot detect Parp protein after total protein isolation Hello everyone; I cannot detect the Parp protein after total protein isolation. I can see my cells die morphologically after a novel...

06 July 2019 9,688 2 View

I can't detect cleaved caspase-3 with western blot?

Hello to everyone. I cannot detect cleaved caspase-3 with western blot in the cells I 48 hours treat with anti-cancer agent. Annexin V and cParp were confirmed by apoptosis. My transfer...

02 March 2019 5,753 6 View

DNA, RNA and protein. What is the ranking of their molecular size?

For example, when RNA is isolated from a cell, how do these molecules (DNA,RNA,protein) pass through the pores?

31 December 2018 1,588 0 View

What do you think about the state of the cell?

HEK-293T is becoming this for a while after being in culture. (20X magnification) Other cells in the culture have no such problem. Can you describe this morphology?

01 January 1970 9,940 3 View

Why is my ASO degraded quickly on agarose gel?

I have been working on Red blood cell-derived extracellular vesicles as Antisense Oligonucleotide (ASO) carriers. We normally run agarose gel to quantify the loading efficiency. I used naked ASO...

06 August 2024 3,130 2 View

Why did the authors extrapolate a phenotype that they experimentally proved in one bacterial strain across the whole genus of the organism?

I aim to be as skeptical as possible regarding whether a pair of orthologous genes results in the same phenotype in their different but related bacterial organisms under similar environmental...

05 August 2024 6,787 4 View

Why my colony PCR results of my recombinant bacterial not showing any results?

I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....

05 August 2024 2,570 3 View

To perform transfection with DharmaFECT Duo in AGS cells. Could you tell me what the ideal concentration is to avoid significant cytotoxicity?

I would like to perform transfection with the reagent DharmaFECT Duo (Horizon) on the AGS cell line. Could you please inform me of the optimal concentration to use without causing cytotoxicity in...

03 August 2024 3,851 1 View

What is the acceptable p-value cutoff for GO enrichment analysis ?

I have an RNA-seq data that I have analysed using Limma-voom and have extracted the gene IDs, log2FC and the p-values. At p value < 0.05, I have over 10,000 DEGs, however, when I run the GO...

31 July 2024 225 2 View

Transfection in HEK293T cells?

Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...

31 July 2024 9,892 4 View

Issues with Protein Expression in Transfected cells?

Hello I am trying to create a stable cell line in HEK293 via Lipofectamine 3000 transfection. My plasmid is a CD63-IL10-GFP construct with Puromycin resistance. I am successful with the...

30 July 2024 6,648 1 View

Inquiry on Maximum Nucleic Acid Volume for 2.5 mL Liposome Solution?

I am currently working on a project involving liposomes and need to determine the maximum volume of siRNA that can be added to a 2.5 mL liposome solution with a total lipid concentration of 10...

30 July 2024 6,420 1 View

How to retain the a GFP tagged gene expression in stable cell line?

Hello , I established a stable cell line expressing GFP tagged to a centrosomal gene having G418 drug selection marker. I validated the stable line by IFA and Western blotting, results are fine....

29 July 2024 5,007 0 View

Does anyone have experience electroporating SKOV3 cells with a large plasmid using Neon transfection?

I have been trying to electroporate SKOV3 cells with a large plasmid (11kb) without much success. Any tips?

29 July 2024 3,229 1 View