Promega Wizard SV 96 Instructions For Use

Wizard ® SV 96 Genomic DNA Purification System
INSTRUCTIONS FOR USE OF PRODUCTS A2370 AND A2371.
Purification of Genomic DNA from Mouse Tail Clippings, Animal Tissues and Tissue Culture
Cells
Preparation of Mouse Tail Clipping and Tissue Samples
1. Cut a 0.5 to 1.2cm mouse tail clipping or weigh up to 20mg tissue sample.
Cut the mouse tail clipping or tissue sample into two pieces and place the
pieces into a 96-well, deep-well plate (not provided).
Digestion Solution
Master Mix
Nuclei Lysis Solution
0.5M EDTA (pH 8.0)
proteinase K, 20mg/ml
RNase A Solution, 4mg/ml
Total Volume
2. Add 275µl of the Digestion Solution Master Mix to each sample. Be sure that
the sample is covered by the solution. Cover the plate with an adhesive seal.
3. Place the plate in a 55°C water bath and incubate overnight (16–18 hours).
Be sure that water does not cover the sample plate. Do not shake.
4. After incubation, dispense 250µl Wizard
Lysates must be warm during processing. Mix the contents by pipetting.
Purification of Genomic DNA from Tail Clippings or Tissue Samples
5. Prepare the vacuum manifold as shown in the figure. Place the Binding Plate
in the Manifold Base. Attach the vacuum line to the vacuum port on the
Manifold Base.
6. Transfer the tissue lysates to the wells of the Binding Plate. Apply vacuum
until all of the lysate has passed through the Binding Plate.
7. Add 1ml of Column Wash Solution (CWA; containing 95% ethanol) to each
well.
8. Apply vacuum until the Column Wash Solution (CWA) passes through the
Binding Plate. Repeat Steps 7 and 8 for a total of 3 washes.
9. After the wells have emptied continue to apply vacuum for an additional
6 minutes to dry the binding matrix.
10. Turn off the vacuum. Release the vacuum line from the Manifold Base and
snap it to the vacuum port in the Vacuum Manifold Collar. Remove the
Binding Plate and blot gently to remove residual ethanol.
11. Place the 96-Well Deep Well Plate in the Manifold Bed and position the
Vacuum Manifold Collar on top. Orient the plate with the numerical column
headers toward the vacuum port.
12. Position the Binding Plate on top of the Manifold Collar. Place the Collar on
top of the Deep Well Plate as shown.
13. Add 250µl Nuclease-Free Water to each well of the Binding Plate and incubate
for 2 minutes at room temperature. (Protocol continued on other side.)
ORDERING / TECHNICAL INFORMATION:
www.promega.com • Phone 608-274-4330 or 800-356-9526 • Fax 608-277-2601
© 2002, 2009, 2012 Promega Corporation. All Rights Reserved.
Volume
per Sample
200µl
50µl
20µl
5µl
275µl
® SV Lysis Buffer into each well.
Quick
P R O T O C O L
Binding Plate
Manifold Base
Vacuum Port
with Insert
in place
B. Washing Apparatus
Binding Plate
Manifold Base
C. Elution Apparatus
Binding Plate
Manifold Collar
Elution Plate
Manifold Bed
Printed in USA. Revised 2/12
Part #9FB070