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1. Where can I find maps and sequences of DUALmembrane or DUALhybrid vectors?
   You can find maps and sequences of all our vectors in the support section of our website."
2. What type of false positives can I expect in the DUALmembrane system?
   Based on our analysis of over 20 screens, We have compiled a preeliminary list of putative false positive interactors. The list can be accessed here. Please keep in mind that this list is subject to...
3. What is a false positive?
   The term 'false positive' means a prey protein which is selected in the yeast two-hybrid system but that does not interact with the bait protein in a physiologically meaningful way. Sometimes, to dist...
4. Can you help me with my yeast transformation?
   You can find a protocol we use for efficient library-scale yeast transformation in our
5. Can you help me with my yeast transformation?
   You can find a protocol we use for efficient library-scale yeast transformation in our
6. Do you maintain a list of common false positives in DUALhybrid screens?
   We do collect data on highly connected proteins, which show up frequently in DUALhybrid screens with different baits and cDNA libraries (false positives). Below is a list of the most frequently found ...
7. How do I construct a bait for a DUALhybrid or DUALmembrane screen?
   For help with constructing baits, preys and libraries in the DUALmembrane system 3, please refer to the DUALmembrane 3 vector selection guide. For help with constructing baits in the DUALhybrid system...
8. Some basic information on replicating plasmids in yeast
   Yeast can be transformed by appropriate plasmids; plasmids used for this purpose are usually constructed as "shuttle vectors" which can be propagated in either Escherichia coli or yeast (Hinnen et ...
9. I have cloned my prey as an x-NubG fusion, but I cannot detect its expression by western blotting.
   Some cDNAs may not be optimally expressed, for instance due to slightly different codon usage in yeast compared to the species of your cDNA's origin. We recommend you use pPR3-STE (DUALmembrane syste...
10. I would like to screen a protein that contains a transmembrane domain. What screen do you recommend?
    Using a DUALmembrane screen you can detect interactions between integral membrane proteins."
11. What does the genotype of your strains mean?
    The genotype of a yeast strain indicates which auxotrophic markers are available for selection and which reporter genes have been integrated into the genome and are available for growth selection or p...
12. Where can I find the complete sequence of DUALmembrane vectors?
    You can find maps and sequences for all our vectors in the support section on our website."
13. I would like to screen a protein that is localized in the membrane. What screen do you recommend?
    Using a DUALmembrane screen you can detect interactions between integral membrane proteins."
14. Can you tell me the genotype of the yeast strain you use in yeast two-hybrid screening?
    The genotype of yeast strain DSY-1 can be found
15. I would like to exclude signal sequences from my bait. What algorithms do you recommend to predict such features of my protein?
    We recommend you visit
16. I would like to exclude transmembrane domains from my bait. What algorithms do you recommend to predict such features of my protein?
    We recommend you visit
17. I would like to screen a protein that contains several transmembrane domain. What screen do you recommend?
    Using a DUALmembrane screen you can detect interactions between integral membrane proteins. We have successfully screened 7-transmembrane domain receptors."
18. I would like to screen a bait protein that is localized in the mitochondria. What screen do you recommend?
    Yeast two-hybrid screens with soluble mitochondrial proteins have been published (see an example here). For mitochondrial membrane proteins we recommend a DUALmembrane screen."
19. I would like to screen a bait protein that is secreted. What screen do you recommend?
    You can remove the signal sequence from your bait and use it in the DUALhybrid system. However, if glycosylation or disulfide formation is crucial for your protein, there may be some difficulties. For...
20. What is the basis of the yeast two-hybrid system?
    An explanation of the yeast two-hybrid system can be found on the