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The Y Chromosome Determines Maleness

The Y Chromosome Determines Maleness

The Y chromosome is a sex chromosome found in several vertebrates and mammals, including humans. In addition to 22 pairs of autosomes, the human males have one X chromosome and one Y chromosome. In these organisms, the presence or absence of the Y chromosome determines the development of male traits.
Evolution
Around 300 million years ago, the two sex chromosomes diverged from two identical autosomal chromosomes. Over time, the Y chromosome has lost most of its genes, shrinking in size. Today,...

Southern Blot

Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...

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Validation of SRY marker for forensic casework analysis.

Vanja Kastelic¹, Bruce Budowle, Katja Drobnic

¹Forensic Science Centre, Ministry of the Interior, Vodovodna 95, Ljubljana, Slovenia. vanja.kastelic@policija.si

Journal of Forensic Sciences

|March 24, 2009

Summary

The amelogenin test for forensic DNA gender determination can fail due to deletions. A validated SRY gene marker offers a reliable alternative, accurately identifying male DNA even in mixed samples.

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Area of Science:

Forensic Science
Molecular Biology
Genetics

Background:

The amelogenin gene assay is standard for forensic DNA gender determination.
Amelogenin assay limitations include deletions, potentially leading to inaccurate gender assignment.
The SRY gene offers a Y-chromosome specific target for gender identification.

Purpose of the Study:

Validate SRY gene primers for specificity, sensitivity, and robustness in forensic applications.
Develop a reliable method for gender determination when amelogenin testing fails.
Assess the performance of the SRY marker in mixed DNA samples.

Main Methods:

Coamplification of an SRY gene marker with short tandem repeat (STR) markers.
Primer validation using specificity, sensitivity, and robustness testing.
Testing with DNA from unrelated Slovenian males and mixed male/female samples.

Main Results:

No null alleles observed in 115 unrelated Slovenian males, indicating high specificity.
Assay demonstrated high sensitivity, detecting as little as 25 pg of template DNA.
No amplification observed with up to 10 ng/µL of female DNA, confirming specificity.
Male DNA detected in mixed samples down to a 1:16 ratio.

Conclusions:

Validated SRY gene primers provide a robust and sensitive method for forensic DNA gender determination.
This SRY marker assay effectively overcomes amelogenin deletion issues.
The validated assay is suitable for identifying male DNA in forensic casework, including mixed samples.