Promega MD1531, Техническое руководство

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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
3
www.promega.com TM248 · Revised 12/18
2. Product Components and Storage Conditions
P R O D U C T S I Z E C AT. #
Y Chromosome Deletion Detection System, Version 2.0 25 reactions MD1531
For Research Use Only. Not for use in diagnostic procedures. The Y Chromosome Deletion Detection System,
Version 2.0, includes:
• 500µl Multiplex A Master Mix
• 500µl Multiplex B Master Mix
• 500µl Multiplex C Master Mix
• 500µl Multiplex D Master Mix
• 500µl Multiplex E Master Mix
• 200u GoTaq
®
DNA Polymerase
• 1ml Nuclease-Free Water
• 90µg 50bp DNA Step Ladder (340ng/µl)
• 2.5µg Male Genomic DNA (50ng/µl)
• 1ml Blue/Orange 6X Loading Dye
Storage Conditions: Store all components at –20°C. Avoid multiple freeze-thaw cycles. The Nuclease-Free Water
may be stored at room temperature (22–25°C).
3. System Requirements
3.A. Template
DNA concentration, purity and size are important considerations to ensure success with the Y Chromosome Deletion
Detection System, Version 2.0. DNA should be free of contaminating protein and salts. The DNA should not be
sheared. Poor-quality DNA may result in increased background or amplification failure. Adding too much or too little
DNA to the reaction can cause amplification failure. Failure can be manifested in several ways; sometimes complete
lack of amplification of all bands is observed with all master mixes, and in other cases dropout of all or portions of the
alleles in individual master mixes will be observed. Several commercially available DNA purification systems, including
the Wizard
®
Genomic DNA Purification Kit (Cat.# A1120, A1125, A1620) and the MagneSil
®
KF, Genomic System
(Cat.# MD1460), produce DNA of sufficient quality for the Y Chromosome Deletion Detection System.
Quantitation of DNA
We recommend quantitating the DNA prior to use in the Y Chromosome Deletion Detection System; either too much or
too little DNA can cause the reactions to fail. Absorbance readings at 260nm can be used to estimate DNA
concentration where 1Au = 50µg of double-stranded DNA/ml. Fifty nanograms of DNA should be used in each
amplification reaction.