Additional file 1

Abstract

Figure S1. A positive-negative selection scheme. The tm246(vps-45) mutants exhibited a temperature-sensitive phenotype. Only tm246 mutants carrying vps-45 rescue transgenes survived at 20°C (positive selection). The tm234(ben-1) mutants were resistant to benzimidazole. tm234 mutants carrying ben-1 rescue transgenes are sensitive and unable to survive on benzimidazole-containing plates (negative selection). Phenotypes of tm234;tm246 were rescued by transgenes, enabling the Ex line to survive at 20°C. The parent Ex animals were treated by UV/TMP, resulting in multi-copy insertion (Is), low-copy Is, and Ex arrays. Because multi-copy Is animals and Ex animals were highly likely to have the ben-1 transgene, only low-copy Is animals carrying the vps-45 and not the ben-1 transgene were selected.

Full text

ben-1(tm234);vps-45(tm246)!
+ ben-1
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ben-1(tm234);vps-45(tm246)

Positive-Negative Selection
ben-1(tm234);vps-45(tm246)!
+ vps-45

ben-1(tm234);vps-45(tm246)!
+ vps-45 & ben-1

20˚C 
15˚C 
Benomyl NGM
Normal NGM
TMP/UV insertion
Ex F1!
Multi-copy Is F1!
Low-copy Is F1
20˚C 
Low-copy Is F1
Normal NGM
20˚C 
Normal NGM
Ex P0
tm234;tm246
Microinjection of selection markers!
+injection marker!
(+plasmid X)
Positive selection
Negative selection
Supplementary Figure1
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