What is the range for the forward primer and reverse primer and how can we identify the best primer?

More Muhammad Sarfaraz Iqbal's questions See All

Why different HCV patients with same genotype respond differently to interferon and Ribaviron combination?

can any explain some genetic differences responsible for EVR,RVR,SVR and NVR

02 March 2015 7,069 9 View

What is minimum requirement of Oxygen pressure for human breathing?

what is the minimum amount of oxygen in Torr for survival

10 November 2014 4,867 2 View

Can we use DNA extracted from serum for Polymorphism analysis?

I have serum samples and want to know if I can use that serum to get DNA for Polymorphism analysis

09 October 2014 1,869 3 View

Can we extract human DNA from Plasma ?

I have plasma samples and want to extract human DNA ,is to possible to extract the DNA from plasma ?

09 October 2014 9,362 20 View

How we can design a best primer for polymorphism investigation?

what are the normal values of parameters used in Primer designing ?

07 August 2014 8,462 2 View

I can't see the ssDNA band after performing asymmetric PCR. Is there any way to do this?

After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...

08 August 2024 1,668 3 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 View

Anyone having idea about VN primer for miRNA primer design ?

How to design VN primer to attach with universal reverse primer

05 August 2024 2,116 3 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

PCR showing no bands - Master mix and primers didn't mix?

Hello everyone, I performed a PCR yesterday, and the results showed no bands on the gel. Of course, I probably missed some crucial steps, like adding my samples to the PCR strips themselves, for...

31 July 2024 2,406 6 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

Do you mix and add all primers together in a singel PCR reaction (4 total primers for 2 mutations) when using Agilent's multi-site mutagenesis kit?

I want to introduce 2 mutations using Agilent's multi-site mutagenesis kit, I designed the primers using their online tool and it gave me 2 sets for primers (1 set for each mutation) so I was...

25 July 2024 6,517 3 View

Can I please ask why my samples from anaerobic bioreactor giving me different size PCR product even after multiple runs?

Hi everyone, I have extracted DNA from a biogas bioreactor using Qiagen kit and prep cDNA library then used this library as template to optimize primers for qPCR (taken from papers). Some of the...

23 July 2024 1,329 5 View

Why my negative control siRNA is decreasing the target gene's expression?

Hi Everyone, I'm using an siRNA kit to knock down a target gene. The kit guarantees that the negative control doesn't target any sequence in mouse genome, and when I use BLAST I don't find any...

23 July 2024 2,673 6 View

Could someone please provide a list of journals that accept the application of methodologies in nature-based solutions?

I am looking for journals that admit the publication of results from applying the IUCN Standard for designing and monitoring Nature-based solutions. Many thanks for considering my request.

23 July 2024 6,444 2 View

Ganapati Bhat

Ideal primer length will be 18-25 bp. Based on this you can enter in NCBI primer blast. Best primer set must have higher score and almost near Tm and GC %. You need to check self complimentary and primer dimer from other tools like FastPCR .

Muhammad Sarfaraz Iqbal

19-24 bp for forward and 18-22bp for reverse