How to purify a special lipid, such as cholesterol and phospholipids?

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 7,035 4 View

Which laptop configuration should i choose for bioinformatics?

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 6,936 0 View

Can SDS-PAGE detect monomer and polymer?

Can SDS-PAGE detect monomer and polymer? If it can, DTT should be added prior to load in gel?

05 June 2018 3,720 1 View

Which concentration of the Bis could the native page need?

05 June 2017 9,348 3 View

What is the white cloudy precipitation after lysing HEK-293T cells?

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 375 0 View

Why CHAPS cannot solve in water after shaking?

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 6,309 0 View

Could native page detect tiny mobility changes?

10 November 2016 8,066 0 View

How much concentration of CHAPS can i use in lysis buffer?

10 November 2016 6,497 3 View

Could I get a high yield human protein production in MDCK?

04 May 2016 4,596 3 View

Western Blot can not detect the expression of protein, although the green fluorescence is expressed. Why ?

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 2,751 4 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 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

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 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

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 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

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

Did anyone ever try using pLVX-TetOne Plasmid with a copGFP as a selection marker instead of Puromycin?

When using a lentiviral vector for inducible expression of genes using the Tet-On system. In the literature, I saw a widely used of pLVX-TetOne-Puro Plasmid, is there a reason why people prefer to...

25 February 2021 1,011 3 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View