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Digital karyotyping (CAT#: STEM-GT-0020-WXH)

Introduction

Digital karyotyping is a technique used to quantify the DNA copy number on a genomic scale. Short sequences of DNA from specific loci all over the genome are isolated and enumerated. This method is also known as virtual karyotyping. Using this technique, it is possible to detect small alterations in the human genome, that cannot be detected through methods employing metaphase chromosomes. Some loci deletions are known to be related to the development of cancer. Such deletions are found through digital karyotyping using the loci associated with cancer development.




Applications

Detecting copy-number changes.
Identifying gene-specific alterations.

Procedure

1. Cleave with mapping enzyme (SacI)
2. Ligate biotinylated SacI linkers to digested DNA
3. Cleave with fragmenting enzyme (NlaIII) and isolate with streptavidin beads
4. Ligate LS linkers to bound genomic DNA
5. Release genomic tags using tagging enzyme (Mmel)
6. Ligate to form ditags
7. PCR amplify, concatenate, sequence tags and map to genome
8. Evaluate tag density

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