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Wizard® Genomic DNA Purification Kit
INSTRUCTIONS FOR USE OF PRODUCTS A1120, A1123, A1125 AND A1620.
Quick
Blood
PROTOCOL
Isolation of Genomic DNA from Whole Blood
Sample Size 300µl 1ml 3ml 10ml Lysis Protein DNA Solution Precipitation Rehydration Nuclei Solution Isopropanol Solution 300µl 100µl 300µl 100µl 1ml 330µl 1ml 150µl 3ml 1ml 3ml 250µl 10ml 3.3ml 10ml 800µl
Cell 900µl 3ml9ml 30ml
Add Cell Lysis Solution. Incubate.
As little as 20µl can be processed with this system. Please see Technical Manual #TM050, Section 3.C.
Centrifuge. Discard supernatant. Vortex pellet. Add Nuclei Lysis Solution. Mix. Add Protein Precipitation Solution. Centrifuge.
Red Blood Cell Lysis 1. Using volumes from the table above, combine the appropriate volumes of Cell Lysis Solutionand blood. Mix by inversion. 2. Incubate for 10 minutes at room temperature. 3. Centrifuge: ≤300µl sample 13,000–16,000 × g* ; 20 seconds 1–10ml sample 2,000 × g ; 10 minutes 4. Discard supernatant. Vortex pellet. Nuclei Lysis and Protein Precipitation 5. Using volumes from the table above, add Nuclei Lysis Solution and mix by inversion. 6. Add Protein Precipitation Solution; vortex for 20seconds. 7. Centrifuge: ≤300µl sample 13,000–16,000 × g* ; 3 minutes 1–10ml sample 2,000 × g ; 10 minutes DNA Precipitation and Rehydration 8. Transfer supernatant to a new tube contaning isopropanol (using volumes from table above). Mix. 9. Centrifuge: ≤300µl sample 13,000–16,000 × g* ; 1 minute 1–10ml sample 2,000 × g ; 1 minute 10. Discard supernatant. Add 70% ethanol (same volume as isopropanol).11. Centrifuge as in Step 9. 12. Aspirate the ethanol and air-dry the pellet (10–15 minutes). 13. Rehydrate the DNA in the appropriate volume of DNA Rehydration Solution for 1 hour at 65°C or overnight at 4°C.
*Maximum speed on a microcentrifuge.
Transfer supernatant to new tube containing isopropanol.
Centrifuge.
Discard supernatant. Add ethanol.
Centrifuge.
2818MA11_9A
Aspirateethanol. Air-dry pellet. Rehydrate DNA.
••••••••••• ••••••• •••
Additional protocol information is available in Technical Manual #TM050, available online at: www.promega.com
ORDERING / TECHNICAL INFORMATION: www.promega.com • Phone 608-274-4330 or 800-356-9526 • Fax 608-277-2601
©1999–2010 Promega Corporation. All Rights Reserved. Printed in USA. Revised 10/10 Part #9FB022
Wizard®Genomic DNA Purification Kit
INSTRUCTIONS FOR USE OF PRODUCTS A1120, A1123, A1125 AND A1620.
Quick
Cells or tissue with Nuclei Lysis Solution.
PROTOCOL
Isolation of Genomic DNA from Animal Tissue and Tissue Culture Cells
Prepare Tissues Tissue Culture Cells: Centrifuge at 13,000–16,000 × g* for 10 seconds. Wash the cell pellet with PBS, vortex and then add 600µl of Nuclei Lysis Solutionand mix by pipetting. Animal Tissue: Add 10–20mg of fresh or thawed tissue to 600µl of chilled Nuclei Lysis Solution and homogenize for 10 seconds. Alternatively, use 10–20mg of ground tissue. Incubate at 65°C for 15–30 minutes. Mouse Tail: Add 600µl of chilled EDTA/Nuclei Lysis Solution to 0.5–1cm of fresh or thawed mouse tail. Add 17.5µl of 20mg/ml Proteinase K and incubate overnight at 55°C withgentle shaking. Lysis and Protein Precipitation 1. Add 3µl of RNase Solution to the cell or animal tissue nuclei lysate and mix. Incubate for 15–30 minutes at 37°C. Cool to room temperature. 2. Add 200µl of Protein Precipitation Solution. Vortex and chill on ice for 5 minutes. 3. Centrifuge at 13,000–16,000 × g* for 4 minutes. DNA Precipitation and Rehydration 4. Transfer supernatant to a freshtube containing 600µl of room temperature isopropanol. 5. Mix gently by inversion. 6. Centrifuge at 13,000–16,000 × g* for 1 minute. 7. Remove supernatant and add 600µl of room temperature 70% ethanol. Mix. 8. Centrifuge as in Step 6. 9. Aspirate the ethanol and air-dry the pellet for 15 minutes. 10. Rehydrate the DNA in 100µl of DNA Rehydration Solution for 1 hour at 65°C or overnight at 4°C....
INSTRUCTIONS FOR USE OF PRODUCTS A1120, A1123, A1125 AND A1620.
Quick
Blood
PROTOCOL
Isolation of Genomic DNA from Whole Blood
Sample Size 300µl 1ml 3ml 10ml Lysis Protein DNA Solution Precipitation Rehydration Nuclei Solution Isopropanol Solution 300µl 100µl 300µl 100µl 1ml 330µl 1ml 150µl 3ml 1ml 3ml 250µl 10ml 3.3ml 10ml 800µl
Cell 900µl 3ml9ml 30ml
Add Cell Lysis Solution. Incubate.
As little as 20µl can be processed with this system. Please see Technical Manual #TM050, Section 3.C.
Centrifuge. Discard supernatant. Vortex pellet. Add Nuclei Lysis Solution. Mix. Add Protein Precipitation Solution. Centrifuge.
Red Blood Cell Lysis 1. Using volumes from the table above, combine the appropriate volumes of Cell Lysis Solutionand blood. Mix by inversion. 2. Incubate for 10 minutes at room temperature. 3. Centrifuge: ≤300µl sample 13,000–16,000 × g* ; 20 seconds 1–10ml sample 2,000 × g ; 10 minutes 4. Discard supernatant. Vortex pellet. Nuclei Lysis and Protein Precipitation 5. Using volumes from the table above, add Nuclei Lysis Solution and mix by inversion. 6. Add Protein Precipitation Solution; vortex for 20seconds. 7. Centrifuge: ≤300µl sample 13,000–16,000 × g* ; 3 minutes 1–10ml sample 2,000 × g ; 10 minutes DNA Precipitation and Rehydration 8. Transfer supernatant to a new tube contaning isopropanol (using volumes from table above). Mix. 9. Centrifuge: ≤300µl sample 13,000–16,000 × g* ; 1 minute 1–10ml sample 2,000 × g ; 1 minute 10. Discard supernatant. Add 70% ethanol (same volume as isopropanol).11. Centrifuge as in Step 9. 12. Aspirate the ethanol and air-dry the pellet (10–15 minutes). 13. Rehydrate the DNA in the appropriate volume of DNA Rehydration Solution for 1 hour at 65°C or overnight at 4°C.
*Maximum speed on a microcentrifuge.
Transfer supernatant to new tube containing isopropanol.
Centrifuge.
Discard supernatant. Add ethanol.
Centrifuge.
2818MA11_9A
Aspirateethanol. Air-dry pellet. Rehydrate DNA.
••••••••••• ••••••• •••
Additional protocol information is available in Technical Manual #TM050, available online at: www.promega.com
ORDERING / TECHNICAL INFORMATION: www.promega.com • Phone 608-274-4330 or 800-356-9526 • Fax 608-277-2601
©1999–2010 Promega Corporation. All Rights Reserved. Printed in USA. Revised 10/10 Part #9FB022
Wizard®Genomic DNA Purification Kit
INSTRUCTIONS FOR USE OF PRODUCTS A1120, A1123, A1125 AND A1620.
Quick
Cells or tissue with Nuclei Lysis Solution.
PROTOCOL
Isolation of Genomic DNA from Animal Tissue and Tissue Culture Cells
Prepare Tissues Tissue Culture Cells: Centrifuge at 13,000–16,000 × g* for 10 seconds. Wash the cell pellet with PBS, vortex and then add 600µl of Nuclei Lysis Solutionand mix by pipetting. Animal Tissue: Add 10–20mg of fresh or thawed tissue to 600µl of chilled Nuclei Lysis Solution and homogenize for 10 seconds. Alternatively, use 10–20mg of ground tissue. Incubate at 65°C for 15–30 minutes. Mouse Tail: Add 600µl of chilled EDTA/Nuclei Lysis Solution to 0.5–1cm of fresh or thawed mouse tail. Add 17.5µl of 20mg/ml Proteinase K and incubate overnight at 55°C withgentle shaking. Lysis and Protein Precipitation 1. Add 3µl of RNase Solution to the cell or animal tissue nuclei lysate and mix. Incubate for 15–30 minutes at 37°C. Cool to room temperature. 2. Add 200µl of Protein Precipitation Solution. Vortex and chill on ice for 5 minutes. 3. Centrifuge at 13,000–16,000 × g* for 4 minutes. DNA Precipitation and Rehydration 4. Transfer supernatant to a freshtube containing 600µl of room temperature isopropanol. 5. Mix gently by inversion. 6. Centrifuge at 13,000–16,000 × g* for 1 minute. 7. Remove supernatant and add 600µl of room temperature 70% ethanol. Mix. 8. Centrifuge as in Step 6. 9. Aspirate the ethanol and air-dry the pellet for 15 minutes. 10. Rehydrate the DNA in 100µl of DNA Rehydration Solution for 1 hour at 65°C or overnight at 4°C....
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