Q1: How do I elute from the included Spin-Column?
The design of the Zymo-Spin V-P Column does make the elution step a little tricky. Sometimes air bubbles can develop while adding the elution buffer to the column, which will result in not being able to add all 400 ul to the column. In order to avoid this, we recommend inserting the tip of the micro pipet directly into the column at an angle until the tip is touching the matrix. Then, slowly expel the elution buffer while removing the tip from the column. Some end users find it easier to use the smaller 200 ul pipet tip and load the column twice with 200 ul using the above technique.
Q2: What is the composition of the ZymoPURE Elution buffer?
10 mM Tris-HCl, 0.1 mM EDTA, pH 8.5.
Q3: Can the ZymoPURE Kits be used with other bacteria?
Yes, please contact technical support for an application note.
Q4: 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.
Q5: Are the ZymoPURE kits compatible with any commercially available vacuum manifold?
Yes, any vacuum that uses standard luer-lock connectors is compatible.
Q6: 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.
Q7: 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.
Q8: Is there a protocol for low-copy number plasmid DNA?
Yes, the low-copy protocol can be found in Appendix A of the kit protocol.
Q9: Can I process more than the recommended volume of bacterial culture or use enriched growth media?
Yes, however, special care should be taken throughout the entire protocol since there is much more biomass. We recommend centrifuging the neutralized lysate before loading onto the supplied syringe filter. For further guidance on dealing with low-copy plasmids, please contact our technical support team. Exceeding the recommended volume for high copy plasmids can cause to overloading and subsequent clogging of columns, which can reduce DNA yield/quality or result in failed preps.
Q10: 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.
