Template:SBB-Protocols Zymo1: Difference between revisions
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===Regular Zymo Cleanup=== | ===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. | 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. | ||
====note: if the product is less than 300bp, add isopropanol==== | |||
#Add 180 uL of Zymo ADB buffer (brown bottle) to a 33uL or 50uL reaction. | #Add 180 uL of Zymo ADB buffer (brown bottle) to a 33uL or 50uL reaction. | ||
Revision as of 10:28, 30 June 2009
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.
note: if the product is less than 300bp, add isopropanol
- 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 PE or Zymo Wash buffer (which is basically 70% ethanol)
- spin through, discard waste.
- Add 200 uL of PE or 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
