What is the difference between Tm (1 M Na+) and Tm (50 mM Na+)?

12 December 2017 0 9K Report

Hello everyone, I received primers, in the certificate of analysis are mentioned 2 Tm, what do they mean and which one should I use? Thank you in advance.

Badges
Science method

More Hossem SAHRAOUI's questions See All

How to prepare (dilute) labeled (marked) primers?

Hello everyone! I would like to know if there is a special way and precautions to follow to dilute lyophilised marked primers (with FAM and HEX)? thanks in advance

01 February 2019 7,832 4 View

Could we publish using XLSTAT software?

I would like to know if the use of some statistical software (such as XLSTAT) is not recommended to publish. Thank you

31 December 2015 8,751 8 View

On what criteria some specific microsatellites were recommended by the FAO and the ISAG for the characterization of animal genetic resources ?

Microsatellites recommended by the FAO and the ISAG for the animal genetic characterization, are they related to characters (qualitative or quantitative)? Or they were chosen on other criteria?...

10 November 2015 3,327 6 View

How do I calculate polypeptides pHi, especially when we have repeated amino acid?

How to calculate pHi of these peptides: 1) Arg-Ala-Arg-Ala-Phe 2) His-Phe-Trp-Asp-Asp thank you

04 May 2015 9,618 3 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

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

Where can I get PCR Primers for COI barcoding of birds?

We are in need of primers for a PCR protocol for chicken genome. We are looking at the COI barcode, 648-bp fragment of the 5' end of COI. Specifically Primers FalcFA, BirdR1, CO1_ExtF,...

16 July 2024 118 2 View