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Table 2.

Generation of Sall4-null ES cells

Vectors
Selection at second roundNumber of coloniesIntegration event
ExperimentFirst roundSecond roundRandomRetargetingSecond allele
pGK Neo pGK Hyg Hyg 134 116 16 
pGK Hyg pGK Neo Neo 115 68 44 3* 
IRES Hyg IRES β-geo Neo 20 12 
 IRES Hyg IRES β-geo Neo+Hyg 13 12 1* 
Vectors
Selection at second roundNumber of coloniesIntegration event
ExperimentFirst roundSecond roundRandomRetargetingSecond allele
pGK Neo pGK Hyg Hyg 134 116 16 
pGK Hyg pGK Neo Neo 115 68 44 3* 
IRES Hyg IRES β-geo Neo 20 12 
 IRES Hyg IRES β-geo Neo+Hyg 13 12 1* 

Two rounds of targeting were carried out to disrupt both the alleles of Sall4 in ES cells.

*

Two out of three clones in Experiment 2 and one clone in Experiment 3 were finally dominated by contaminating heterozygous cells.

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