No title

Particularly, I want to know the cultured cell ATP assay Protocol, and data analysis (I am working on GBM cell line). Any prompt reply would be highly appreciated Thank you,

09 October 2018 2,182 0 View

No title

Particularly, I want to know the cultured cell ATP assay Protocol, and data analysis (I am working on GBM cell line). Thank you,

09 October 2018 6,230 0 View

What is the required concentration of all three (Packaging, Envelop, and Transfer) vectors in Lentivirus Packaging?

I need to package lentivirus to generate gene of interest stable brain tumor cell line so, I want to know what is the required concentration of packaging, envelop and transfer vectors to transfect...

31 December 2017 2,649 2 View

How to interpret JC-10 Staining results

Hello Dear Researchers does anyone have experience to interpret JC-10 staining result for Mitochondria Membrane Potential. Could someone please explain me in detail with images because yet i don't...

11 December 2016 8,266 0 View

Manual Protein Extraction Protocol?

10 November 2016 8,427 2 View

What if re-autoclave PBS twice for Cell culture use?

what if re-autoclave PBS twice for Cell culture use? it will work normal or not?

09 October 2016 1,465 0 View

What should be the minimum DMSO concentration to dissolve Temozolomide drug?

09 October 2016 2,937 3 View

Not getting desired band of inserts after digesting with respective enzymes?

I want to do subcloning of two genes (gene X and gene Y) in pcDNA 3.1+ expression vector. Gene X is having KpnI and NheI restriction site, gene Y having BamHI and NheI restriction sites....

08 September 2013 3,434 12 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 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

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

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

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View