Q1: What is the composition of the ZymoPURE™ Elution buffer?
10 mM Tris-HCl, 0.1 mM EDTA, pH 8.5.
Q2: I ran out of ZymoPURE™ Wash 2. Can I substitute it with a homemade solution or Wash Buffer from another kit?
No, the ZymoPURE™ kits are only compatible with ZymoPURE™ Wash 2 and we cannot disclose a substitution due to the sensitive nature of the recipe. Additional ZymoPURE™ Wash 2 can be purchased separately.
Q3: What type of vacuum pump do you recommend?
The vacuum pump should be a single or double-staged unit capable of producing at least 400 mm Hg pressure at the vacuum manifold. If less pressure is applied, centrifuge the column prior to washing to remove any residual lysate/buffer remaining in the matrix.
Q4: Are the ZymoPURE kits compatible with any commercially available vacuum manifold?
Yes, any vacuum that uses standard luer-lock connectors is compatible.
Q5: Can an in-house vacuum line be used with the EZ Vac Vacuum Manifold?
Yes, however, the pressure needs to be around 400 mm Hg. Users should take caution, as the pressure of an in-house vacuum line can fluctuate drastically or be significantly reduced, depending on the demand in the building.
Q6: Can the ZymoPURE Kits be used with other bacteria?
Yes, please contact technical support for an application note.
Q7: Is there a protocol for low-copy number plasmid DNA?
Yes, we do have an application note for low-copy number plasmid available. Please contact technical support for a copy.
Q8: Can I process more than the recommended volume of bacterial culture or use enriched growth media?
Yes, however, more attention is required during the lysis and steps. We recommend centrifuging/pelleting the neutralized lysate before loading onto the supplied syringe/bottle-top filter. For high copy plasmids using more culture can cause overloading and subsequent clogging of columns, which can result in poor DNA yield and/or quality.
Q9: Can the ZymoPURE kits be used to isolate large plasmid constructs (BAC/PAC)?
Yes, the standard ZymoPURE protocol has been successfully tested with constructs up to 200 kb. To increase elution of large plasmid DNA, we recommend pre-warming the ZymoPURE Elution Buffer (50 ºC) and increasing the incubation time on column up to 10 minutes prior to centrifugation.
