Is it valid to directly compare expression levels of two genes with different amplicon size?

16 June 2025 0 3K Report

I want to compare the expression of two genes (Gene A: 300 bp amplicon, Gene B: 200 bp amplicon) using SYBR Green qPCR. Is it valid to directly compare their expression levels given the difference in amplicon size?

Badges
Science method

More Sisira Surendran's questions See All

Implementation of interaction integral for XFEM of functionally graded materials from Kim and Paulino (2005). What is wrong in my implementation ?

The equation 49 is of the non-equilibrium formulation which is easier since it needs only the first order derivatives. However, my results don't match with benchmark results. In fact it is close...

24 February 2023 985 0 View

What are the advantages of using C-rate, rather than A/g, to measure battery performance?

Some journal reviewers prefer that the rate performance of batteries be expressed in C rate rather than A/g. However, since battery capacities vary across different chemistries, wouldn't A/g be a...

20 February 2023 973 1 View

I am looking for Maslach burnout inventory - General Survey (MBI-GS) ?

Can anyone share it with me

20 March 2022 2,855 0 View

I am looking for questionnare on Occupational stress, Job satisfaction, Employee burnout and employee productivity for software professionals?

Can anyone suggest some questionnaires on these? Also the questionnaires for each variables must be closer to 20 or so. I would like to keep it short

19 March 2022 1,068 3 View

Is NLDFT pore size distribution calculated via N2 adsorption isotherms at liquid nitrogen temperature reliable to probe pores less than 0.7nm?

In general, pores less than 0.7nm is not probed using N2 gas at liquid nitrogen temperatures. CO2 is the preferred gas to probe the ultra micropores. But, the NDLFT calculations in Micromeritics 3...

15 July 2021 1,872 0 View

Does the adsorbed water or gases affect the CHNS analysis of high surface area porous carbon ?

Adsorbed water and gases can affect the weight of the porous carbon. Some times 6-7% weight loss can occur during the degassing of the sample. Will this weight loss affect the calculation of C,...

08 November 2020 5,598 3 View

How to generate load spectrum in fe safe?

I wanna know the procedure to generate loading and load spectrum?

01 January 2019 825 3 View

How to do welding simulation in ansys workbench ?

I want to do Co2 Mig Welding in pipe.Is it is possible to use ansys workbench for welding simulations.(Without using Plugins)

31 August 2018 5,161 6 View

How to apply bolt load in abaqus explicit?

i have run bolt load in abaqus implicit/standard then i imported this output to explicit but deformation created due to bolt load is not showing so i needed to apply bolt load in explicit but i...

08 August 2018 6,658 1 View

How can I get DPPH IC 50 value ?

For doing DPPH free radical scavenging assay, I have used ascorbic acid as the standard. and took different concentrations (5,10,15,20,25 and 30 microgram/ml) for both standard as well as for...

15 July 2018 6,407 12 View

I'm optimizing a tetra-ARMS PCR protocol with amplicon sizes: 165bp, 123bp and 93bp. Why, in the gel, only 165bp is visible while I'm expecting all 3?

It's an end-point PCR protocol. I'm using 1.5% agarose gel with SyBR Safe dye and TBE as a running buffer, visualization on BioRad XR+ system. I was primarily thinking of primer efficiency,...

01 August 2024 4,673 4 View

How to retain amplicon yield after Ampure bead cleanup, following a 2-stage PCR protocol?

Hello all, I have been trying to follow a 2-stage PCR protocol used to amplify barcodes of a large yeast library, as per Nyugen et al. (2022) -...

30 July 2024 841 2 View

Are there always been barcodes, apapters and primer sequences in the FASTQ files of NGS?

Hello researchers, Sorry for my stupid question. I am learning the QIIME2 workflow for analyzing some 16s amplicon NGS fastq data. I found a very nice paper with data and code public available...

20 July 2024 5,405 2 View

Why I am not getting exact amplicon size after PCR?

I have synthesized a gene specific primer for amplification of my target gene. However, I am getting amplicon of only 100 bp rather than 210 bp (target amplicon size). So far I have tested so many...

05 June 2024 1,587 7 View

Why digestion like band appear in pcr product?

my gene of interest size is about 4kb. I used hi fidelity taq pol as well as Q5 from NEB but still its happening. my forward primer composed of a 4 adenosine then restriction site followed by 6x...

05 May 2024 4,558 2 View

Regarding primers annealing temperature ?

I recently designed the qrtpcr primers from the 3'utr of the plant's genomic DNA keeping in mind that the annealing temperature should be 60 degree. Once I received the primers first to optimise...

25 April 2024 5,935 3 View

Help on Modified Nanopore Sequencing to do Sanger-like sequencing for PCR amplicon from bisulfite converted DNA?

Hi, I'm planning to do sequencing using Nanopore. The main idea is to substitute Sanger Sequencing that usually used to sequence PCR amplicon from BCD (Bisulfite Converted DNA) using nanopore...

23 April 2024 6,637 0 View

Why could amplicon sizes appear bigger than they should on agaose gel electrophoresis?

I did PCR of 6 amplicons and when I did agarose gel electrophoresis, they all appear consistently bigger than they should be (e.g. the amplicon should be 4000 bp, but is around band of 5000 bp...

09 April 2024 4,487 5 View

Can SIMPER (vegan, R) analysis be done on binary distance matrices, particularly those containing ASV data?

Hello! I am trying to characterize communities of eukaryotes living in biofilms using V4 region 18s amplicon sequencing data. Working with this type of data is new to me, and very interesting!...

04 April 2024 6,603 1 View

How to start the trials to obtain amplification in my PCR?

Hello, I am currently attempting to obtain amplifications of SoxC from a species of onychophoran belonging to the Peripatidae family using primers designed from a sequence of its sister family,...

20 March 2024 4,626 4 View