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18 June 2019 7,409 8 View

Has anyone else seen a reversal of solutions during an mRNA extraction?

I am trying to do an mRNA extraction to be used in qPCR. I am using trizol. When I add the chloroform, I should see a clear aqueous solution containing my mRNA at the top and the pink phenol...

10 April 2019 7,821 3 View

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22 October 2018 3,303 3 View

Does anyone have a Nocodazole release protocol for A549 cells?

I am doing an experiment to investigate the effect of our compound on cell cycle. I will be using Nocodazole to synchronize my A549 cells but see a wide variety of doses when searching the...

01 January 2018 5,973 3 View

How long does it generally take to recieve the full-text article/journal?

I have made several requests this afternoon for full-text articles/Journals. I am a new user to this site and am just curious how long it generally takes to process those requests. Thank you!

11 December 2017 4,893 5 View

How do you reconstruct a 3d image from a single hologram?

Hi there! I have a confusion on the idea of what a 3D image really is from the hologram reconstruction. Sometimes the paper talks about a 3D volumetric image from a stack of amplitude intensity...

22 October 2016 9,709 3 View

Why do most cell lysis buffers for western have either NP40 OR Triton-X? Is there a reason that you can't mix the two into one universal buffer?

11 April 2016 6,568 2 View

In qPCR, if your target gene dCT is negative can you still use this data as the RQ value will still be positive?

06 April 2016 4,093 3 View

Has anyone noticed poor agreement between replicates using the qiagen one-step RT-PCR kit?

I have been using the kit above for some time now, which performs cDNA conversion and normal qPCR in the same tube. I am noticing that despite using recently calibrated pipettes and preparing my...

30 April 2015 5,024 10 View

What should I do with my primer optimization?

I am optimizing a 16s primer pair and cannot get rid of primer dimers/ nonspecific binding, despite them apparently having no capacity to form primer dimers according to oligoevaluator. They were...

27 June 2014 917 9 View

How can I measure MMP-1 protein activity or protein level from liver tissue?

Instead of checking only the mRNA level, I want to check the active protein level of MMP-1 in Liver tissue from mice. How can I do that?

03 March 2021 1,763 2 View

Does any one know how i can prepare liver tissue with transmission electron microscope with good fixation?

i have problem in preparation of liver tissue with bad cell organelles and how i can adjust osmolarity

03 March 2021 8,890 1 View

IHC on mouse liver hepatocytes?

Can anyone help me choose primary antibodies for mouse liver hepatocytes and also in conjunction Cd8 cells. Also Cd3 cells. I want to look at hepatocytes and adjacent immune cells in mice with...

01 March 2021 6,953 1 View

How to avoid ulcers in s.c. injected tumors ?

Recently, we have injected s.c. B16F10 tumor cells in BL/6 mice. At around 12-15th day some animals showed ulcerations in tumors , with a big hole in the middle (it looks like a volcano). We have...

01 March 2021 9,938 3 View

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 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

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View

No amplification data for qPCR on MxPro?

I have set up a qPCR run on MxPro with SYBR green and ROX as a reference, however, when I go to the analysis section the program seems to only collect data for the dissociation curves and not the...

25 February 2021 8,039 1 View

Looking for genomic DNA extraction kits from mouse tissue (centrifuge-based, not spin-column)?

Hello. I am looking to extract genomic DNA from mouse brain and lungs and I need a kit that is not spin-column based because I need to scale up the solution amounts for 400mg tissue (most cap at...

25 February 2021 155 2 View

What are the pros and cons for mice versus rats?

Hi Everyone, I am designing a pilot study using rodents to assess the effects of simulated space environments (radiation + altered gravity) on microbiome and immune system. I have limited...

24 February 2021 1,825 3 View