Biomod/2013/Harvard/protocols: Difference between revisions
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# Thaw chemically competent cells on ice. | # Thaw chemically competent cells on ice. | ||
# Transfer 50 ul of competent cells to a 1.5 ml microcentrifuge tube (if necessary). | |||
[[Biomod/2013/Harvard/protocols/miniprep | miniprep]] | [[Biomod/2013/Harvard/protocols/miniprep | miniprep]] |
Revision as of 13:03, 17 June 2013
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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.
- 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
- Load the mixture into a Zymo-Spin Column in a Collection Tube.
- Centrifuge at full speed (≥10,000 x g) for 30 seconds. Discard flow through.
- 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.
- 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.
Bacterial Cell Culture Solutions
BLACaM Assays
Chemically Competent Cells Transformation
- Thaw chemically competent cells on ice.
- Transfer 50 ul of competent cells to a 1.5 ml microcentrifuge tube (if necessary).