Wizard Genomic DNA Purification Kit Protocol - Department of ...

Wizard ® Genomic DNA
Purification Kit
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Technical Manual. Please contact Promega Technical Services if you have questions on use
of this system. E-mail: techserv@promega.com.
1. Description..........................................................................................................1
2. Product Components and Storage Conditions ............................................2
3. Protocols for Genomic DNA Isolation ..........................................................5
A. Isolating Genomic DNA from Whole Blood
(300µl or 3ml Sample Volume)...........................................................................5
B. Isolating Genomic DNA from Whole Blood
(10ml Sample Volume) ........................................................................................7
C. Isolating Genomic DNA from Whole Blood
(96-Well Plate).......................................................................................................9
D. Isolating Genomic DNA from Tissue Culture Cells and
Animal Tissue .....................................................................................................11
E. Isolating Genomic DNA from Plant Tissue ...................................................13
F. Isolating Genomic DNA from Yeast................................................................14
G. Isolating Genomic DNA from Gram Positive and
Gram Negative Bacteria ....................................................................................16
4. Troubleshooting...............................................................................................17
5. References .........................................................................................................18
6. Appendix ...........................................................................................................19
A. Composition of Buffers and Solutions ............................................................19
B. Related Products.................................................................................................19
1. Description
The Wizard ® Genomic DNA Purification Kit is designed for isolation of DNA
from white blood cells (Sections 3.A, B and C), tissue culture cells and animal
tissue (Section 3.D), plant tissue (Section 3.E), yeast (Section 3.F), and Gram
positive and Gram negative bacteria (Section 3.G). Table 1 lists the typical yield
for DNA purified from each of these sources.
The Wizard ® Genomic DNA Purification Kit is based on a four-step process (1).
The first step in the purification procedure lyses the cells and the nuclei. For
isolation of DNA from white blood cells, this step involves lysis of the red
blood cells in the Cell Lysis Solution, followed by lysis of the white blood cells
and their nuclei in the Nuclei Lysis Solution. An RNase digestion step may be
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Part# TM050
Revised 12/10 Page 1