EvaGreen® Dye

PCR Quantitative real-time PCR - Bacterial DNA

Experiment
PCR Quantitative real-time PCR - Bacterial DNA
Product
EvaGreen® Dye from Biotium
Manufacturer
Biotium

Protocol tips

Upstream tips
Amplicon size should be consistent for each target sequence and limited to approximately 65 - 100 bp.
Protocol tips
PCR plates, tubes and pipette tips should be UV sterilized for 20-30 mins

Publication protocol

PCR assays for detecting fungi
To detect fungi, the LightCycler Nano (Roche Applied Science) and PCR-clean tubes were used as described above. During the PCR, the PCR reaction mixture (20 μL) contained 2 μL of DNA template or 2 μL (8.0 ng/μL) of DNA extracted from Candida albicans as a positive control, or distilled water (NAKALAI TESQUE, INC.) as a negative control in 50 mM KCl, 2.5 mM MgCl2, 10 mM Tris-HCl (pH 8.3), 200 μM of each deoxynucleoside triphosphate (dNTP), 0.25 μM each of Fungal universal primer, 1×EvaGreen (Biotium Inc.), and 2.0 units (0.4 μL) of conventional thermostable DNA polymerase (r-Taq: Toyobo, Osaka, Japan) supplemented with stock buffer solution. Each sample was incubated for three minutes at 95°C, then denatured for 10 seconds at 95°C, annealed for 15 seconds at 57°C, extended for 20 seconds at 72°C and subjected to fluorescence acquisition for two seconds at 82°C for 40 cycles.

Full paper   Login or join for free to view the full paper.

Reviews

EvaGreen® Dye from Biotium has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Discussion

4 years ago

Author: Germany

What is the optimal concentration for primers in qPCR?

What is the optimal concentration for primers in qPCR? My total volume is 20μl per reaction.

Share your thoughts or question with experts in your field by adding a discussion!

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing PCR Quantitative real-time PCR - Bacterial DNA using EvaGreen® Dye from Biotium.

Paper title
Eukaryote-Made Thermostable DNA Polymerase Enables Rapid PCR-Based Detection of Mycoplasma, Ureaplasma and Other Bacteria in the Amniotic Fluid of Preterm Labor Cases
Full paper
Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Biotium for EvaGreen® Dye below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing PCR Quantitative real-time PCR - Bacterial DNA using EvaGreen® Dye from Biotium. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms