Does PCR once standardized using rat liver differ for another animal? And if yes why is that so?

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Why i do get black particles in cell culture and i also have dead cells adhered to my grown live cells and its like just waves?

I grow LC-540 cells and my cells are clear when fully grown image 1, but when I trypsinize the same plate I see black particles image 2. When I subculture them on a new plate I don't see those...

02 March 2020 6,467 1 View

Help me Identify the problem with these cells (LC540 Leydig cells)?

I grow LC-540 Leydig cells, the problem with me is as shown in the picture a. The ones rounded in yellow circles are black particles attached to the plate loosely it does not show any movement,...

03 April 2019 5,164 5 View

Oocyte swelling assay detail protocol. Aquaporin inhibition studies. Any suggestions for other studies?

I am completely unaware of swelling assay as i am from a different back ground. I have a compound which i hypothesize that it will inhibit aquaporin. I am not sure that it would make any gene or...

02 March 2018 2,823 0 View

Does high or low glucose has its effect on total AMPK (not phosphorylated AMPK) in cells during treatment?

I have been using endothelial cells with various conc of glucose from 1.5mM to 30mM. I have changes in total AMPK protein expression using AMPK alpha1/2 santa cruz. I just went through some...

04 May 2016 8,357 2 View

Best compound or drug to inhibit aquaporin expression other than mercury?

I have been working on aquaporins and literaturre says merrcury inhibits AQPs. Few published articles say Thrombin, TPA, bumetadine, Propofol pholoretin etc.. CAn someone suggest me the best among...

02 March 2016 7,310 8 View

This picture is LC540 leydig cells... The red arrows indicate black dots and the yellow arrows indicates bubbles. What are these stuff?

I'm new to cell culture and i have started with Lc540 leydig cells i started growing the cells and i notice these black dots as shown in the picture in red arrows and it doesn't affect my culture...

08 September 2015 3,987 8 View

Any one working in the field of reproduction with sertoli cells?

Sertoli cell line

10 November 2014 6,501 1 View

How do you mimic diabetes conditions in-vitro to study the effect on reproductive organs (e.g. in testes)?

Diabetes has many complications, one of which is on reproduction (infertility). How can I mimic diabetes condition in a testes cell line? I have seen papers in which a pancreas (beta cell line)...

02 March 2014 7,622 7 View

Why are wistar rats specifically used in diabetic models?

I have also seen in many articles using SD rats, but mostly wistar rats are used is there.

01 February 2014 3,313 2 View

Why are my rats dying after 48 hrs when I induce them with STZ 65?

Rats 0f 250-300mg each mg/kg.

01 February 2014 645 14 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