What is the best way to do inter assay and intra assay validation of an assay?

More Saima Qayum's questions See All

Doing NEXT GENERATION SEQUENCING, What is the minimum required reads of the targeted mutation in order to label a sample mutation positive?

I am doing targeted next generation sequencing targeting a number of somatic mutations. One of the mutations is a point mutation. Now in a sample if I have total 1000 reads, 10 reads are point...

09 October 2019 3,149 1 View

Does product size affect realtime PCR optimization?

I am optimizing a realtime PCR using probes with FAM and TAMRA. But I do not get any results. The product size is 77 basepairs. Is this small product aize the cause of failed optimization?what...

04 May 2019 8,565 4 View

Can we make a stock of forward and reverse primers in a single tube?

I intend to do next generation sequencing for total six mutations in each sample. It involves 6 primer sets. In order to reduce the pipetting time can I make a stock of all primer sets in a single...

01 February 2019 9,367 4 View

Can i use Ambion nuclease free water Not DEPC treated for reconstitution of my primers and probes?

I intend to do a real time PCR using probes( FAM TAMRA). Can I use Ambion Nuclease free water Not DEPC treated to reconstitute my primers and probes?

01 February 2019 9,976 5 View

Dear all, I was wondering if anyone knows whether MGLtools and Autodock can run on Mac OS?

I have tried to download the 64-bit version of MGLtools however, as many times as I have downloaded and uninstall the programs, an error arises that the app needs to be updated. Thank you.

03 March 2021 5,997 2 View

What is your perception on using visual analysis or quantitative approaches to evaluate data from single-case research designs (n of 1 trials)?

Results of single-case research designs (i.e., n-of-1 trials) are often evaluated by visually inspecting the time-series graph and computing quantitative indices. A question our research team is...

03 March 2021 687 1 View

I got 02 moderator values defining Johnson-Neyman significance regions. Is it an error?

Hi everyone, I am conducting research for my Master's thesis. I am using PROCESS by Johnson-Neyman to analyze my Moderator model. I test the relationship between Public Service Motivation and...

03 March 2021 2,350 2 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

Do I need to do linear regression before running mediation analysis using PROCESS MACRO by Hayes ?

I just wanted to check if I need to run a linear regression separately if I am using PROCESS MACRO to run mediation analysis. Thank you.

02 March 2021 4,359 3 View

How to quantitative analize the data of TNBS assay for gelatin?

Estemeed colleagues, I found some issues regarding the quantification of the data for TNBS assay. There are different protocols on how to perform that but it is clear to me the "fil rouge" that...

02 March 2021 2,616 1 View

Do I need to strip the PVDF membrane before staining it with colloidal gold?

Dear Colleagues, After running Western blot on PVDF membrane and detection using ECL, I would like to stain my PVDF with colloidal gold to be able to allign the ECL image with total proteins on...

02 March 2021 7,829 3 View

PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 View

What SPSS analysis should I run with one IV (with two levels) and two DVs?

Hi, I am trying to figure out which test would be most appropriate to run for the following question: Is there a difference in task blood pressure between smokers and non-smokers? There are two...

01 March 2021 8,235 2 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

Engelbert Buxbaum

The following document may be helpful: http://academy.gmp-compliance.org/guidemgr/files/UCM070107.PDF

Karen Hunt

For questtion 1 yes the same sanple is what’s needed if your checking repeatability of the instrument. Different samples will test repeatability of the samples so it depends on what test your doing. For 2 and 3 the minimum is 16 but make sure you validate every part of the process, Engelbert’s suggestion to read is good. Hope this helps

Ako Rezaei

Hi! I've attached a PDF file that describes how to calculate inter and intra-assay CVs are and how to interpret them.