[ GeneKin^® Y-STR Systems ]

Contents

Description
  A. GeneKin^® Y-STR systems (Silver Stain Detection)
  B. Advantages of GeneKin^®  Y-STR Systems typing
 
GeneKin^® Y-STR systems (Silver Stain Detection)

Amplification
  A. Choice of Thermal Cycling Protocol
  B. Amplification Set-Up
  C. Amplification Thermal Cycling
  D. Agarose Gel Electrophoresis of Amplification Products (optional)

Electrophoresis and Detection
 Polyacrylamide Gel Preparation
  A. Notes
  B. Procedure

 Polyacrylamide Gel Electrophoresis
  A. Gel Pre-Run
  B. Sample Preparation
  C. Sample Loading
  D. Gel Electrophoresis

Silver Staining
  A. Notes
  B. Procedure

Results

References

Description

A. GeneKin^® Y-STR systems (Silver Stain Detection)

  Interspersed short tandem repeat (STR) loci consist of tandem arrays raging from 1 to 6 base pairs in length (1-4). These STR sequences represent a significant fraction of eukaryote (e.g., human) genomes and are detected mostly by polymerase chain reaction (PCR) analysis (5-8). Alleles of these loci are differentiated by the number of copies of the repeat sequence contained within the amplified region and are distinguished from one another using silver stain following electrophoretic separation.

  The usefulness of Y-chromosome markers for studies of human population genetics and forensic/paternity analysis has recently  been recognized (9-10). The human Y-chromosome of the non-recombining portions has special features: a haploid transmission pattern; and father-to-son transmission. The DNA sequence of the non-recombining portions contains a genetic record only of the mutational events that occurred in their past (11-17).

  GeneKin^® Y-STR Systems are more useful for investigating and reconstructing the male mediated forensic/paternity testing (18). Moreover, GeneKin^® Y-STR Systems will be of great benefit in analyzing mixed DNA samples, in investigating sexual assaults, as well as in paternity testing (paternal kinship test) where the alleged father is not available but other patrilineal relatives are (19).

B. Advantages of GeneKin^® Y-STR Systems typing

  The GeneKin Y-STR Systems typing provide a rapid, non-radioactive method, which can be used to evaluate small amounts (e.g., 50 ng) of human DNA. The protocols detailed in this manual describe the use of silver staining (20) to detect the presence of amplified STR products following their separation by denaturing polyacrylamide gel electrophoresis.

  In addition to these advantages, the Y-STR loci chosen for inclusion in the GeneKin^® Y-STR Systems contain alleles of discrete and separable lengths. This allows the construction of allelic ladders, which contain fragments of the same lengths as several or all known alleles for the locus. Visual comparison between the allelic ladder and amplified samples of the same locus allows rapid and precise assignment of alleles.

 
GeneKin^® Y-STR systems (Silver Stain Detection)

  The GeneKin^® Y-STR Systems provide all of the component required to perform 100 amplification reactions except Taq DNA polymerase and sample DNA.

Storage Conditions: Store all components at -20¡É. The pre- and post-amplification components (Allelic Ladder Mix, Loading Solution and Control DNA) are sealed in separate packages to prevent contamination. These systems are guaranteed for at least 6 months from date of purchase.

Table 1. GeneKin^®  Y-STR Systems Information^*