Template:SBB-Protocols Zymo2: Difference between revisions
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===Small-Frag Zymo Cleanup=== | ===Small-Frag Zymo Cleanup=== | ||
The following procedure removes the polymerase, dNTPs, buffer, and most of the oligonucleotides from a PCR reaction for fragments smaller than 300bp. It also will remove the buffer and restriction enzymes from a restriction digest reaction. | |||
#Add 100 uL of Zymo ADB buffer (brown bottle) to the reaction. | |||
#Transfer into the Zymo column (small clear guys) | |||
#Add 500uL of Ethanol and pipette up and down to mix | |||
#spin through (15s), discard waste. | |||
#Add 200 uL of Zymo Wash Buffer (which is basically 70% ethanol) | |||
#spin through (15s), discard waste. | |||
#Add 200 uL of Zymo Wash Buffer | |||
#spin through, discard waste. | |||
#spin for 90 seconds, full speed to dry. | |||
#elute with water (spin 60s) into a fresh Eppendorf tube | |||
Latest revision as of 09:10, 2 February 2012
Small-Frag Zymo Cleanup
The following procedure removes the polymerase, dNTPs, buffer, and most of the oligonucleotides from a PCR reaction for fragments smaller than 300bp. It also will remove the buffer and restriction enzymes from a restriction digest reaction.
- Add 100 uL of Zymo ADB buffer (brown bottle) to the reaction.
- Transfer into the Zymo column (small clear guys)
- Add 500uL of Ethanol and pipette up and down to mix
- spin through (15s), discard waste.
- Add 200 uL of Zymo Wash Buffer (which is basically 70% ethanol)
- spin through (15s), discard waste.
- Add 200 uL of Zymo Wash Buffer
- spin through, discard waste.
- spin for 90 seconds, full speed to dry.
- elute with water (spin 60s) into a fresh Eppendorf tube
