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Catalog 2007

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GenePack DNA PCR for cancer mutation detection tests provide simple and convenient analysis of one specific mutation.

All reagents are lyophilized and stabilized ...

..... for a convenient and simple usage, storage and transportation.

Each kit contains 100 tubes with lyophilised Master Mix with the following components:

• Taq DNA polymerase plus heat-sensitive Taq DNA inhibitors

• 4 primers for allele-specific PCR

• dNTP’s

• special salts and stabilizers

• loading dye (bromphenol blue)

• Separate Tube of dilution buffer

The Master-Mix can be transported stored at ambient temperature (+15-30 °C)  for more than 1 Year without any change in performance of mutation detection.
To start PCR one have to add genomic DNA, dilution buffer and water.  After PCR the product will be loaded on the Agarose gel (no need for loading dye).
The size of the fragments will indicate the presence or absence of the mutation in the sample studied.
 

Introduction cancer mutation detection tests

The most popular types of familial cancers are familial breast cancer and breast-ovarian cancer syndrome. Approximately 1% of all women are strongly genetically predisposed to the breast/ovarian cancer.
Women who are carrier of the  mutation of BRCA1 or BRCA2 genes will get cancer with the probability more than 80% in the life span.
Analysis of  mutations in these genes provides the possibility for early tumor detection in mutations carriers. The search for cancer mutation in the particular patient is complex due to the variety of mutations described and due to the involvement of multiple genes in predisposition formation.
Meanwhile, some mutations in BRCA1 and BRCA2 genes are recognized as the most frequent mutations in familial breast/ovarian cancers. Analysis of these mutations in cancer patients can easily reveal mutation carriers even without knowledge of familial history of the patient.

Principle of mutation detection tests

Each kit is designed for the detection of one specific mutation. The mutation detection is based on tetra primer allele-specific PCR. Flanking primers provide amplification of a region around possible mutation in all cases. Internal primers can discriminate mutation allele and wild-type allele by the corresponding amplicon size (Figure 1):

To start PCR one have to add genomic DNA, dilution buffer and water.  After PCR the product will be loaded on the Agarose gel (no need for loading dye).
The size of the fragments will indicate the presence or absence of the mutation in the sample studied. Fig. 2 shows an example of 5382insC mutation detection in BRCA1 gene:

                                             Figure 2.

Analysis of DNA sample on the presence of 5382insC mutations in BRCA1 gene.
1,2,3 – No mutation
            detected
4 – Mutation 5382insC
      (heterozygote)
5 - Molecular Weight
     Marker

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