Is anyone willing to share pancreatic cancer cell lines like MIA PaCa-2 and PANC-1 or any other ?

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Is fold change true representative of differential gene expression?

Hi Everyone, I have a simple query. When we estimate differential gene expression between two data sets or two conditions, we generally observed log(base 2)fold change. However, the fold change...

09 October 2018 2,776 4 View

Regarding fluorescence microscopy ?

Hi Guys, I am trying to make a stable cell line from NIH/3T3 by transfecting with my plasmid of interest. After transfection, while I was changing the medium, I applied a bit harsh pressure on...

09 October 2018 1,556 3 View

What are locus specific primers? How are they different from normal primers?

Hi everyone! I cam across term called "locus specific primer". I came across this term in 3C (chromatin conformation capture) method. I tried to google about it but could not understand much. Are...

06 July 2018 8,665 1 View

Do anyone know about an offline tool for RNA seq analysis?

Hello everyone I have to travel a lot in next few days but I don't want to waste time. Does anyone know any tool which can be used to perform RNA seq data analysis offline (from fastq files). I...

06 July 2018 4,575 4 View

Why it is important to take equal amount of total RNA for reverse transcription reaction (cDNA preparation)?

Hey everyone, I am working on liver tissue samples. I have FACS sorted 3 populations from same tissue and I want to study the gene expression between these populations using qPCR. One of the...

05 June 2018 6,435 11 View

What are the two population clouds in FACS data?

Dear all, I am trying to determine the DNA content in cell population by Hoechst 33342 DNA specific stain using flow cytometry analysis (BD FACS fortessa). I am observing two different clouds (...

04 May 2018 6,407 4 View

Karyotyping protocol for AML-12 cell lines?

Hi everyone, I was wondering if anyone have done the karyotyping for AML-12 cell lines (adult mouse liver cell lines)? I believe the protocol for karyotyping should be same like any other adherent...

31 December 2017 503 1 View

Regarding fluorescence microscopy ?

Hi Everyone, I have used Phalloidin to label F-actin in ethanol fixed cells ( I know formaldehyde might have been a better choice for fixation). I am new to new to this and I am experiencing that...

10 November 2017 9,387 2 View

Culturing conditions for HepG2?

Hello All, I just started using HepG2 cell lines and I would like to know the culturing conditions. I have read through the ATCC guidelines for this cell line and they recommend EMEM+10% FBS....

09 October 2017 4,308 5 View

Is anyone using formaldehyde to fix cells and extract RNA from these cells later?

I want to sort cells using FACS, followed by fixation of sorted populations using formaldehyde or ethanol. I would like to extract total RNA from these sorted and fixed cells later. Has anyone...

07 August 2017 6,459 2 View

Why Do TDS and EC Increase with Larger Wastewater Volumes, While BOD and COD Decrease?

I have carried out MFC experiments on three different volumes, 50, 500 and 1000 mL of wastewater. Results after MFC treatment shows that TDS and EC are more in larger volumes of water i.e. TDS and...

09 August 2024 9,621 0 View

Could it be a cell culture contamination?

Hello everyone! I observed in my culture (htert-RPE1 cells) an orange- red particle at the bottom of the dish. It is visible to the naked eye as a very very small red dot. Could it be a...

09 August 2024 2,824 3 View

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

How to normalize and take the significance of the MTT OD values with 3 replicates for the same cell-line?

Hi, I have a question about normalizing the MTT OD values for doing the statistical analysis. So, if we have 3 different plates and we call them 3 different replicates, so, first we would...

07 August 2024 8,106 4 View

Why activated CAR-Jurkat cell could not kill targets?

Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...

06 August 2024 641 2 View

How much total RNA concentration to be extracted from sorted plasma cells from bone marrow of C57BL/6 mice for RT-PCR ?

i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...

05 August 2024 8,835 1 View

Weak DAPI staining after immunohistochemistry - how to improve?

After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...

05 August 2024 9,637 2 View

The Curse of Evolution and Complexity?

Brain and body mass together are positively correlated with lifespan (Hofman 1993). The duration of neural development is one of the best predictors of brain size, and conception is the best...

05 August 2024 6,247 3 View

How to isolate lymphocytes from mouse spleen?

I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...

04 August 2024 9,913 7 View

How to Add Missing Water Molecules in Protein-Membrane Simulations?

I have protein-membrane simulations (PDB, PSF, DCD) and have noticed that water molecules near the protein are not visible in the simulations. How can I fix this issue? Is there a way to place the...

04 August 2024 1,200 2 View