Have anybody faced repeated contamination of AGS cell line?

Hi I have AGS cell lines in culture for our experiments. Recently, I started facing fungal/ yeast contamination of the AGS cell line in culture continuously, once they are 90% confluent. The...

11 December 2018 1,395 11 View

How can I synchronize the liver cells?

01 February 2017 1,404 5 View

What is the role of hydrocortisone in the culture of MCF 10A cells?

Can anybody explain the role of hydrocortisone for the growth of MCF 10A cells? Is it necessary for the culture of the cells in DMEM/F12 HAM medium? Can I culture the cells without hydrocortisone?

04 May 2016 7,979 0 View

What are the steps for flask coating?

I have to use pre-coated flasks for some of my cell lines (BEAS-2B, THLE-2). Can anybody help me with the flask coating (BSA, collagen and fibronectin)? For how long can I store the prepared...

04 May 2016 4,069 3 View

I embedded the samples for TEM in a day old resin and can't pellet it. Is there a way to save my samples?

I'm preparing the samples for my experiment for TEM. I did the resin embedding step. After adding the pure resin I centrifuged the cells at 1,500 rpm 5 minutes but could't pellet it well. The...

01 February 2016 873 4 View

Is it a good practice to leave the samples in gluteraldehyde fixative over a few days and continue processing for electron microscopy?

I want to leave the samples for electron microscopy in gluteraldehyde fixative over the weekend and continue the process next week. Will it affect the samples? How long can the samples be left in...

01 February 2016 9,746 6 View

How to determine the concentration of nanoparticles to be used in experiments?

I want to know how to determine the concentration / amount of nanoparticles that should be used for the experiment

31 December 2015 7,963 11 View

What is the procedure for extraction and estimation of total lipids from cultured animal cells?

11 December 2015 3,560 3 View

How do I increase the yield of my PCR product?

Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...

02 March 2021 4,029 3 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

How to address this issue in the analysis of Real-Time PCR results?

To dear Researchers, I was analyzing a series of concentration for estimation of Real-Time PCR efficiency. The concentration was 1:10. I used MS-excel to evaluate Slope. The result of slope was -8...

01 March 2021 8,683 4 View

No title

Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

01 March 2021 5,311 1 View

MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 View

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 View

Can thermal cycler machines be problematic during PCR?

Hi, my question is about the heating of thermal cycler machines and I hope some of you had experienced a similar thing previously. There are two thermal cycler machines in the lab(BioRad) and for...

26 February 2021 4,777 4 View

Calculating volume of DNA required from DNA concentration?

Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...

26 February 2021 5,029 2 View

Has anyone used the Illumina ITS1 primer sets?

Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...

25 February 2021 6,969 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