Difference between revisions of "Zymo Research Clean and Concentrate for Plasmid DNA"
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3. Transfer mixture to Zymo-SpinTM Column in a Collection Tube. | 3. Transfer mixture to Zymo-SpinTM Column in a Collection Tube. | ||
| − | 4. Centrifuge for 30 seconds and discard the flow through. | + | 4. Centrifuge for 30 seconds (11,000 - 16,000 rpm) and discard the flow through. |
5. Add 200µl of DNA Wash Buffer to the column. Centrifuge for 30 seconds. | 5. Add 200µl of DNA Wash Buffer to the column. Centrifuge for 30 seconds. | ||
Latest revision as of 18:25, 18 June 2018
Clean and Concentrate Protocol
Ratios (DNA Binding Buffer : Sample)
2:1 plasmid, genomic DNA
5:1 PCR product, DNA fragment
7:1 ssDNA
When using plasmid DNA that has been through Miniprep Protocol.
1. Combine 2 volumes of DNA Binding Buffer to each volume of plasmid. (Ex. 200µl DNA Binding Buffer : 100µl plasmid DNA)
2. Mix gently by vortex.
3. Transfer mixture to Zymo-SpinTM Column in a Collection Tube.
4. Centrifuge for 30 seconds (11,000 - 16,000 rpm) and discard the flow through.
5. Add 200µl of DNA Wash Buffer to the column. Centrifuge for 30 seconds.
6. REPEAT Step 5.
7. Add 25µl of dH20 (55̊C) to the column. Let sit at room temperature for approximately 2 minutes.
8. Transfer column to 1.5ml microcentrifuge tube. Centrifuge for 30 seconds.
Results:
Start: 65.5ng/µl
After Clean and Concentrate: 50ng/µl
Yield: 76.3%
