Type-it HRM PCR Kit (2000)

High-resolution melting (HRM) analysis Bacterial

Experiment
High-resolution melting (HRM) analysis Bacterial
Product
Type-it HRM PCR Kit (2000) from Qiagen
Manufacturer
Qiagen

Protocol tips

Protocol tips
DNA quantification prior to performing the HRM procedure is a necessary step to avoid introducing error due to modifications in the shape of the melt curves

Publication protocol

Singleplex PCR amplifications: Singleplex endpoint PCRs of ST131 DNA template and individual MLST primer sets were performed using the Qiagen Type-it HRM PCR kit in a final volume of 25 μl with 10 ng of DNA template and 12.5 pMol each primer (30). DNA quantification prior to performing the HRM procedure is a necessary step to avoid introducing error due to modifications in the shape of the melt curves. The cycling conditions consisted of a 5-min denaturation at 95°C, followed by 30 cycles of a 10-s denaturation at 95°C and 30 s of annealing at 51°C and then by 10 s of extension at 72°C. Amplicons were analyzed by HRM at a 0.1°C resolution from 50 to 95°C to determine the absolute melt curve of each amplicon. The derivative melt curves of MLST amplicons were compared to the melt curves predicted by uMelt software, developed at the University of Utah, to confirm sequence identity. PCR amplicons were also separated on a 1% agarose gel and visualized with ethidium bromide to confirm that only a single product of the expected size was amplified.

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

Reviews

Type-it HRM PCR Kit (2000) from Qiagen has not yet been reviewed for this experiment

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

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing High-resolution melting (HRM) analysis Bacterial using Type-it HRM PCR Kit (2000) from Qiagen.

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

Manufacturer protocol

Download the product protocol from Qiagen for Type-it HRM PCR Kit (2000) below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing High-resolution melting (HRM) analysis Bacterial using Type-it HRM PCR Kit (2000) from Qiagen. 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