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14. Apply vacuum until the Nuclease-Free Water passes through the Binding Plate.
15. Repeat Steps 13 and 14 for a total elution volume of 500µl.
16. Release the vacuum and remove the Binding Plate. Remove the Manifold
Collar, making sure the Deep Well Plate remains positioned in the Manifold
Bed. If droplets are present on the top of the wells, gently tap the plate on
the bench top. Eluate volumes may vary but are generally 400–450µl. Store
samples at –20 or –70°C by covering the plate tightly with a plate sealer.
Purification of Genomic DNA from Tissue Culture Cells
1. Wash the cells once with sterile 1X PBS.
2. Add 150µl of Wizard
®
SV Lysis Buffer to the washed cells. Pipet to mix.
3. 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.
4. Transfer the cell lysates to the wells of the Binding Plate. Apply vacuum until
all lysate has passed through the wells of the Binding Plate.
5. Add 1ml of Column Wash Solution (CWA; containing 95% ethanol) to each
well.
6. Apply vacuum until the Column Wash Solution (CWA) has passed through the
Binding Plate. Repeat Steps 5 and 6 for a total of 3 washes.
7. After the wells have emptied, continue to apply vacuum for an additional
6 minutes to dry the binding matrix.
8. 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.
9. 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.
10. Position the Binding Plate on top of the Manifold Collar. Place the Collar on
top of the Deep Well Plate as shown.
11. Add 250µl Nuclease-Free Water to each well of the Binding Plate and incubate
for 2 minutes at room temperature.
12. Apply vacuum for 1 minute.
13. Release the vacuum and remove the Binding Plate. Remove the Manifold
Collar, making sure the Deep Well Plate remains positioned in the Manifold
Bed. If droplets are present on the top of the wells, gently tap the plate on the
bench top. Eluate volumes are generally 225µl. Store samples at –20 or
–70°C by covering the plate tightly with a plate sealer.
See additional protocol information in Technical Bulletin #TB303, available online at:
www.promega.com/protocols
Purification of Genomic DNA from Mouse Tail Clippings, Animal Tissues or Tissue Culture
Cells (continued)
Wizard
®
SV 96 Genomic DNA Purification System
INSTRUCTIONS FOR USE OF PRODUCTS A2370 AND A2371.
P R O T O C O L
Quick
Manifold Base
Manifold Base
Manifold Bed
Manifold Collar
Binding Plate
Binding Plate
A. Genomic DNA Binding Apparatus
B. Washing Apparatus
C. Elution Apparatus
Binding Plate
Vacuum Port
with Insert
in place
Elution Plate
2626MB03_1A
© 2002, 2009, 2012 Promega Corporation. All Rights Reserved.
Printed in USA. Revised 2/12
Part #9FB070
