Could cells from proliferation express the plasmid which i have transfected using Lipofectamine2000?
What about the mechanism?
How to purify a special lipid, such as cholesterol and phospholipids? I need to purify the whole cholesterol in the blood to the exclusion of other lipids. Is there any special molecules to purify...
04 May 2019 7,140 4 View
which laptop configuration should i choose for bioinformatics? I'm going to study the bioinformatics. But, the professor told me that my laptop isn't suitable for bioinformatics. Therefore, i...
11 December 2018 7,112 0 View
Can SDS-PAGE detect monomer and polymer? If it can, DTT should be added prior to load in gel?
05 June 2018 3,830 1 View
The PI of two proteins are 4.96 and .4.86 and the MW of them are both roughly 15KD. Which concentration of the Bis could the native page need? How about the voltage when running?
05 June 2017 9,422 3 View
What is the white cloudy precipitation after lysing HEK-293T cells? My lysis buffer is composed of 1%NP-40, 150mMNaCl, 50mMTris, 5mMEDTA. After lysing 30min and centrifuge at 1,2000 *g with 30min,...
04 May 2017 469 0 View
I added 1 g CHAPS into 100ml water and shook it. However, it didn't become clear. Why? How could i do to obtain a clear 1% CHAPS?
10 November 2016 6,389 0 View
My protein is 25KD and its pI is 5.5. How much concentration of my native page could be? The native page could deter the tiny mobility changes? And which is the right running buffer, 0.5*TBE or...
10 November 2016 8,133 0 View
How much concentration of CHAPS can i use in lysis buffer? I would like to purify protein. But only CHAPS is compatible with mass spectrometry.
10 November 2016 6,631 3 View
Can I get a high yield human protein production in Madin Darby Canine Kidney(MDCK)? Which factor should be focused on if I choose a cell line to yield human protein production? Why people choose...
04 May 2016 4,670 3 View
Western Blot can not detect the expression of protein, although the green fluorescence is expressed. Why ? The vector is CMV promoter and the sequence is that eGFP-2A site-Myc-insert. There are...
03 April 2016 2,826 4 View
I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...
08 August 2024 4,645 2 View
Hi all, I need to introduce an ARS (autonomously replicating sequence) in my plasmid but I'm not sure which position would be the best. Does anyone have any suggestion? A picture of the plasmid...
05 August 2024 1,573 4 View
I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...
05 August 2024 9,059 3 View
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
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
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
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
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
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
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