I want to...

Automate Throughput	Decrease Labor Prep Time
Reduce Input gDNA	Stop Using Manual Slab Gels
Analzye Larger Fragments	Eliminate Clean Up Step
Increase Throughput	Reduce Time To Results
Eliminate Labeled Primers

I want to Eliminate Clean Up Step

Time- consuming bottlenecks - caused by clean-up steps and post heteroduplex DNA enzyme digestion, are a major drawback to the traditional method for mutation detection. Bottlenecks occur because of the requirement to add EDTA to the heteroduplex DNA:enzyme mixture, which in turn necessitates removal of the EDTA in order to satisfactorily separate and visualize the cleaved DNA. This clean-up step not only adds time but the need for additional costly reagent (Sephadex and specialized plates) and reagent preparation.

By following the Mutation Detection Kit Protocol and using the AdvanCE™ FS Nucleic Acids Analyzer, clean-up steps are eliminated. Instead, a simple reagent addition step is added that can be automated by liquid handling instruments. The purpose of the diluent buffer is to reduce the enzyme activity while at the same time adjusting the salts to appropriate levels for efficient DNA injection onto the AdvanCE™ FS Nucleic Acids Analyzer system.

1. Add 2µl PCR product to 2µl enzyme solution
2. Spin 10 sec
3. Incubate @45C for 45'
4. Add 24µ diluent buffer
5. Place on ice until CE

One of the added benefits of the AdvanCE™ FS Nucleic Acids Analyzer process is that samples can be prepared and placed on ice prior to analysis, allowing samples to be batch processed, further improving workflow and collectively reducing labor per sample.

The following factors allow the elimination of the sample clean up steps on the AdvanCE™ FS Nucleic Acids Analyzer:

1. Streamlined process that utilizes only 3 pipeting steps prior to electrophoresis.
2. High detection sensitivity and efficient cleavage of heteroduplex DNA.
3. Limited reaction volumes.