Plasmid Flashcards
Is it okay if I left my P1 buffer at room temperature?
Yes, P1 buffer is fairly stable at room temperature. For long-term use, I would recommend storing at 4C
Can the ZymoPURE kits be used with other bacteria?
Yes, the standard protocol should work with other gram-negative bacteria. However, the user will need to validate the protocol for their particular species. For gram-positive bacteria, please refer to the Gram-Positive Bacteria Protocol in the Appendix of the kit protocol.
Can I substitute ____ from another kit or a homemade method?
No, the ZymoPURE buffer and column system is unique and cannot be interchanged with other Zymo products or home brew methods
What is the recommended OD600 for my culture?
0.2-0.35
I have a low copy plasmid, are there any modifications I need to make?
Yes, we have a low copy plasmid protocol in the appendix
Can I use TB or another enriched media?
Yes, however, special care should be taken throughout the entire protocol since there is more biomass. We recommend increasing the incubation time during lysis and centrifuging the neutralized lysate before loading onto the supplied syringe filter. Exceeding the recommended culture volume or using enriched media for high copy plasmids may overload and subsequently clog the columns, which can reduce DNA yield/quality or result in failed preps.
Does the ZymoPURE Midiprep, Maxiprep or Gigaprep remove Endotoxins?
Yes, all three of these kits contain an Endotoxin removal column after elution. The final Endotoxin level should be below 0.025EU/ug
Does the ZymoPURE Miniprep remove Endotoxins?
No, the ZymoPURE Miniprep does not contain Endotoxin removal. Our larger size ZymoPURE kits do contain Endotoxin removal columns.
Which vacuum manifold should I use?
You can use any vacuum manifold with standard luer-lock connectors and a vacuum pressure of at least 400mmHg. We hsell a vacuum manifold (catalog S7000).
How long should I incubate my culture for?
Incubation times depend on strain, culture conditions, and antibiotic used. We recommend culturing until the OD measures 0.2-0.35
My ZymoPURE Binding Buffer and/or ZymoPURE P2 has precipitate in it. Are they still okay to use?
Yes, sometimes the buffers precipitate during shipping. You can heat them at 30-37C for 10 minutes and mix by inversion. Do not microwave.
I got very low yield with the ZymoPURE kit. Can you help me troubleshoot?
Low yield is usually caused by overgrown culture or exceeding kit limitations. Please answer the questions below so we can get a better idea of what could be causing the low yield:
troubleshooting Qs
What is the difference between the regular ZymoPURE kits and the ZymoPURE Express?
The ZymoPURE Express uses a direct lysis method by skipping the resuspension step in P1 buffer.
Can I use an antibiotic other than Ampicillin with the Mix & Go Transformation Kit & Buffer Set?
Yes, other antibiotics besides Ampicillin can be used with this kit, but they require an outgrowth step. Please see the protocol for step-by-step information.
What is the composition of the ZymoPURE Elution buffer?
10 mM Tris-HCl, 0.1 mM EDTA, pH 8.5.
