Template:SBB-Protocols Zymo2: Difference between revisions
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#Add 500uL of Ethanol and pipette up and down to mix | #Add 500uL of Ethanol and pipette up and down to mix | ||
#spin through (15s), discard waste. | #spin through (15s), discard waste. | ||
#Add 200 uL of | #Add 200 uL of A4 Wash Buffer (which is basically 70% ethanol) | ||
#spin through (15s), discard waste. | #spin through (15s), discard waste. | ||
#Add 200 uL of | #Add 200 uL of A4 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 (spin 60s) into a fresh Eppendorf tube | #elute with water (spin 60s) into a fresh Eppendorf tube |
Revision as of 21:14, 21 February 2011
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 A4 Wash Buffer (which is basically 70% ethanol)
- spin through (15s), discard waste.
- Add 200 uL of A4 Wash Buffer
- spin through, discard waste.
- spin for 90 seconds, full speed to dry.
- elute with water (spin 60s) into a fresh Eppendorf tube