Template:SBB-Protocols Zymo1: Difference between revisions
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#Transfer into the Zymo column (small clear guys) | #Transfer into the Zymo column (small clear guys) | ||
#spin through, discard waste. | #spin through, discard waste. | ||
#Add 200 uL of | #Add 200 uL of Zymo Wash Buffer (which is basically 70% ethanol) | ||
#spin through, discard waste. | #spin through, discard waste. | ||
#Add 200 uL of | #Add 200 uL of Zymo Wash Buffer | ||
#spin through, discard waste. | #spin through, discard waste. | ||
#spin for 90 seconds, full speed to dry. | #spin for 90 seconds, full speed to dry. | ||
#elute with water into a fresh Eppendorf tube, use the same volume of water as the volume of the original reaction | #elute with water into a fresh Eppendorf tube, use the same volume of water as the volume of the original reaction |
Latest revision as of 09:09, 2 February 2012
Regular Zymo Cleanup
The following procedure removes the polymerase, dNTPs, buffer, and most of the oligonucleotides from a PCR reaction. It also will remove the buffer and restriction enzymes from a restriction digest reaction.
- Add 180 uL of Zymo ADB buffer (brown bottle) to a 33uL or 50uL reaction.
- Transfer into the Zymo column (small clear guys)
- spin through, discard waste.
- Add 200 uL of Zymo Wash Buffer (which is basically 70% ethanol)
- spin through, discard waste.
- Add 200 uL of Zymo Wash Buffer
- spin through, discard waste.
- spin for 90 seconds, full speed to dry.
- elute with water into a fresh Eppendorf tube, use the same volume of water as the volume of the original reaction