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Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516
TM248 · Revised 12/18
www.promega.com
1. Description
The Y Chromosome Deletion Detection System, Version 2.0
(a,b)
, is intended for research use only and provides a rapid
method for the detection of specific regions of the human Y chromosome. This system is designed to detect deletions
occurring in YqAZF. Further mapping of common deletion breakpoints relative to palindromes 1 through 8 may be
performed in additional experiments (1). This system consists of 20 primer pairs that are homologous to previously
identified and mapped sequence-tagged sites (STS; 2–6). This product is not intended for use in diagnostic
applications.
These primers will amplify nonpolymorphic short DNA segments from the Y chromosome when used in polymerase
chain reactions (PCR; 1). The primers have been combined into five sets for use in multiplex PCR. This makes it
possible to determine the presence or absence of all 20 sequence-tagged sites by performing five parallel PCR
amplifications.
Four of the Multiplex Master Mix sets (Multiplexes A–D) contain a control primer pair that amplifies a fragment of the
X-linked
SMCX
locus. The fifth Multiplex Master Mix set, Multiplex E, contains a control primer pair that amplifies a
unique region in both male and female DNA
(ZFX/ZFY
). These control primer pairs are internal controls for the
amplification reaction and the integrity of the genomic DNA sample. Finally, Multiplex E Master Mix includes a primer
pair that amplifies a region of the
SRY
gene.
Figure 1 shows a typical amplification of male genomic DNA, as well as a negative DNA control, for each of the five
Multiplex Master Mixes.
Figure 1. Multiplex analysis of wildtype male genomic DNA.
Lane 1 shows the amplification products from
reactions using Multiplex A Master Mix with male DNA, and lane 2 shows the no-DNA control. Lane 3 shows the
amplification products from reactions using Multiplex B Master Mix with male DNA, and lane 4 shows the no-DNA
control. Lane 5 shows the amplification products from reactions using Multiplex C Master Mix with male DNA, and
lane 6 shows the no-DNA control. Lane 7 shows the amplification products from reactions using Multiplex D Master
Mix with male DNA, and lane 8 shows the no-DNA control. Lane 9 shows the amplification products from reactions
using Multiplex E Master Mix with male DNA, and lane 10 shows the no-DNA control. Lanes M contain the 50bp DNA
Step Ladder (Cat.# G4521). The reactions were performed on a Perkin-Elmer Model 480 thermal cycler. The gel is a
4% NuSieve
®
3:1 Plus TBE Buffer Reliant
®
gel.
4322TA09_3A
M
50 –
200 –
300 –
400 –
500 –
bp
100 –
– 50
– 200
– 300
– 400
– 500
– 100
A
1 2 M
B
3 4 M
C
5 6 M
D
7 8 M
M
E
9 10
