Transfection should I grow the cells in Media without antibiotic and FBS ?

Transfection should i grow the cells in Media without antibiotic and FBS ?

05 June 2015 9,263 5 View

What should i expect on my SDS gel after analyzing my immunoprecipitation experiment?

05 June 2015 2,941 0 View

Please help I am new in Plasmid construction ?

04 May 2015 3,281 3 View

How can the cell attach to cell culture plate?

How can the cell attach to cell culture plate? I know there are two reasons. First, electrostatic reaction second, extracellular matrix but need more explanation.

03 April 2015 6,757 4 View

Can anyone suggest why the growth of my cell culture of TNBC MBA-MD-436 is slow?

03 April 2015 4,256 3 View

Need help I just start my first cell culture experiment .what's rounded shape cells means in cell culture during the growing in media no trypsin ?

03 April 2015 2,174 7 View

Can anyone please help me to find information about these 6 cell lines ?

kuramochi -ovcar - ovarsha- HEYA8- SKVO3- OVAMANA Please help me to learn more about these cell lines.

10 November 2014 624 2 View

Why am I getting a very dark high background in WB Kifc1 antibody?

I'm working now on western blot and trying to detect Kifc1, but it gives me a dark background and I washed for 2 hrs and used 5% BSA-TBST blocking. The same problem of high background is very dark.

10 November 2014 281 5 View

What's the importance of making standard curve for protein estimation ?

i do it but i can't understand the mechanism and how we can estimation our protein using Bradford soln ?

10 November 2014 4,244 4 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 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

What is the difference between genotype and haplotype data in the ped format?

Hi everybody In the ped format for genotype, alleles of any SNP are represented by two columns (one for each allele, separated by a space). Is a column sufficient for the haplotype to...

27 February 2021 1,965 1 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

What's the concentration recommend for recombinant protein use in western blotting?

I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...

24 February 2021 8,514 3 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

IF troubleshooting: looking at an intracellular protein that localizes to the cell's and how to reduce increase in signal due to cell ruffles?

I am currently trying to stain for PI3KBeta in MDA-MB-231 cells. My current issue is that 1) beta stains all over the cell and 2) there is an increase in signal at the edge of the cell and this...

24 February 2021 8,582 1 View

Is it efficient to do liquid biopsy to investigate the gene expressions of the tumor?

To do liquid biopsy and look for specific gene expressions there are so many published papers. Thera are different protocols and procedures used to isolate circulating cell free RNA or mRNA from...

23 February 2021 7,326 5 View

QPCR reaction efficiency with odd results what does this indicate?

I am working on a CFX 96 machine to determine reaction efficiency of my gene of interest. After performing a serial dilution of my cDNA template, I was able to find great reaction efficiencies for...

23 February 2021 809 3 View

Could not amplify CD28 and CD3z ectodomain from cDNA of Jurkat Cells ?

For my experiments, I have to express ectodomains of human CD28 and CD3z. As a source, I extracted RNA from Jurkat cells and verified the quality of RNA on the gel. I, then, prepared cDNA using...

23 February 2021 7,051 4 View