Biomod/2013/Harvard/protocols: Difference between revisions
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* For ssDNA purification, add 7 volumes | * For ssDNA purification, add 7 volumes | ||
2. | 2. Load the mixture into a Zymo-Spin Column in a Collection Tube. | ||
3. Centrifuge at full speed (≥10,000 x g) for 30 seconds. Discard flow through. | 3. Centrifuge at full speed (≥10,000 x g) for 30 seconds. Discard flow through. | ||
4. Add 200 µl of DNA Wash Buffer (from kit) to the column and centrifuge for 30 seconds. Repeat the wash step, this time centrifuging for 1 minute. | 4. Add 200 µl of DNA Wash Buffer (from kit) to the column and centrifuge for 30 seconds. Repeat the wash step, this time centrifuging for 1 minute. | ||
5. Place the Zymo-Spin Column into a new 1.5 ml tube. Add ≥6 µl of DNA Elution Buffer (from kit) or water directly to the column matrix and spin to elute the DNA. | 5. Place the Zymo-Spin Column into a new 1.5 ml tube. Add ≥6 µl of DNA Elution Buffer (from kit) or water directly to the column matrix and spin to elute the DNA. | ||
Revision as of 12:59, 17 June 2013
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HARVARD
BIODESIGN
BIOMOD 2013
-
<html><a href="http://openwetware.org/wiki/Biomod/2013/Harvard"><li>HOME</li></a></html>
<html><a href="http://openwetware.org/wiki/Biomod/2013/Harvard/introduction"><li>INTRODUCTION</li></a></html>
<html><a href="http://openwetware.org/wiki/Biomod/2013/Harvard/design"><li>DESIGN</li></a></html>
<html><a href="http://openwetware.org/wiki/Biomod/2013/Harvard/methods"><li>METHODS</li></a></html>
<html><a href="http://openwetware.org/wiki/Biomod/2013/Harvard/protocols"><li>PROTOCOLS</li></a></html>
<html><a href="http://openwetware.org/wiki/Biomod/2013/Harvard/team"><li>TEAM</li></a></html>
<html><a href="http://openwetware.org/wiki/Biomod/2013/Harvard/lab"><li>LAB NOTEBOOK</li></a></html>
<html><a href="http://openwetware.org/wiki/Biomod/2013/Harvard/results"><li>RESULTS</li></a></html>
<html><a href="http://openwetware.org/wiki/Biomod/2013/Harvard/references"><li>REFERENCES</li></a></html>
Protocols
DNA Clean and Concentrate
This protocol is used to clean and concentrate PRC products after DPN1 digestion. We use the ZYMO DNA Clean & Concentrator - 5 Kit for this protocol.
1. Add 2-7 Volumes of DNA Binding Buffer (from kit) to each volume of DNA sample.
- For plasmid or genomic DNA 2kb, add 2 volumes.
- For PCR or short DNA fragments, add 5 volumes.
- For ssDNA purification, add 7 volumes
2. Load the mixture into a Zymo-Spin Column in a Collection Tube.
3. Centrifuge at full speed (≥10,000 x g) for 30 seconds. Discard flow through.
4. Add 200 µl of DNA Wash Buffer (from kit) to the column and centrifuge for 30 seconds. Repeat the wash step, this time centrifuging for 1 minute.
5. Place the Zymo-Spin Column into a new 1.5 ml tube. Add ≥6 µl of DNA Elution Buffer (from kit) or water directly to the column matrix and spin to elute the DNA.
