Validating NGS data with regular Taqman or advanced Taqman miRNA assay?

Hi, I wish to validate NGS data of miRNA sequencing from human plasma samples. The obvious way to do it is of course Taqman, but there are two different assays: the 'traditional' one in which...

31 December 2018 6,963 0 View

How to find reference genes for qPCR from RNA-seq data?

Hi everyone, I did RNA-seq for small RNA, and now I want to confirm the results with qPCR. What is the best way to find reference miRNA genes from the NGS data, for the normalization of the...

31 December 2018 712 4 View

Discrepancy between DESeq2 data and raw data?

Hi, i performed NGS for small RNA and ran DESeq on the data. there is one miRNA that is upregulated by 2-fold in the patients compared to healthy controls. However, when i look at the raw...

06 July 2018 3,398 3 View

Protocol for plasma preparation?

Hi, I need to isolate plasma from blood for RNA extraction afterwards. Does it have to be in a cold centrifuge? Do you need in general to isolate plasma in a refirigerated centrifuge or is it...

04 May 2018 5,998 5 View

Doing more PCR cycles to get higher concentrations of libraries?

Hi, I am preparing small RNA libraries for NGS, using the QIAseq kit of Qiagen Usually I get Canadian concentrations of around 1-10 ng/ul, but in the last batch all of the libraries were

04 May 2018 1,975 5 View

Is "camel hump" in tape station a sign for adapter dimer?

09 October 2017 2,433 1 View

Perfusion or post-fixation?

Hi, I have brains that express GFP under a promoter of an endogenous protein so they actually express a fusion protein. I want to visualize GFP and thus know where the endogenous protein expresses...

04 May 2017 3,628 6 View

PCR plates or PCR strips for library prep?

Hello, If I use a PCR plate for library prep and I need the products after each step in the protocol (in my case it's polyadenylation, RT and PCR), is it safe to open the plastic foil that covers...

03 April 2017 9,111 3 View

Working with PCR plates for large-scale preparation of cDNA libraries for sequencing?

Hi, Does anyone have experience with preparing cDNA libraries from small RNA, for next-generation sequencing? If so, has anyone tried doing it in PCR plates instead of strip tubes? I have a large...

01 January 1970 9,534 3 View

Any recommendation for a bioinformatic pipeline for small RNA sequencing?

01 January 1970 4,046 4 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

Which of the Covid-19 Vaccines would elicit the best immunoprotection and why?

I want to know which of the available Covid-19 Vaccines would elicit the best immunological response and why

02 March 2021 1,388 3 View

How does the gut microbiome affect the fibrotic process within the liver?

and Can the gut microbe be used for the treatment or diagnosis of liver fibrosis?

02 March 2021 557 2 View

Sample dilution for quantitative analysis using ELISA?

If the detection range is in ng/ml but the reference range is in ug/ml for a molecule or protein in serum or plasma .how to dilute and what is the initial volume to be taken for quantitative analysis

02 March 2021 7,670 3 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

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

How to Eliminate cosine error and abbe error in LASER measurement system(for example Rhenishaw XL80)?

I like to do project in laser measurement system. My objective is to eliminate or reduce the cosine error, abbe error in Laser measurement system. I have Rhenishaw XL80 system. Please tell the...

28 February 2021 4,890 3 View

How to measure angle between laser travel path and machine axis in Laser Measurement system?

25 February 2021 1,987 3 View

Experimental Design: How deal with pseudreplicates with immunological and clinical-chemical paramaeters?

Hello everybody, I am investigating the influence of a toxinon various clinical-chemical and haematological parameters in fish. I have three treatments (control, low dose, high dose) in my...

24 February 2021 1,243 1 View