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Figure 3 | BMC Genetics

Figure 3

From: Essential and non-essential DNA replication genes in the model halophilic Archaeon, Halobacterium sp. NRC-1

Figure 3

PCR assay to screen for knockout alleles of Halobacterium sp. NRC-1 orc genes. Lanes 1–20 contain products obtained from individual PCR reactions using total genomic DNA extracted from 20 individual Foar colonies as template for each gene examined respectively, M denotes DNA ladder. A. Extrachromosomal orc genes. Primers residing ~1000 bp 5' and 1000 bp 3' to each orc gene (orc2, orc3, orc4, orc5, or orc9) in Halobacterium sp. NRC-1 were used with total genomic DNA from individual colony isolates in PCR reactions to screen for orc gene knockouts. For orc2, orc3, orc4, orc5, and orc9, knockout alleles where obtained are ~2000 bp in size, while wild-type alleles are approximately 800, 300, 1200, 1400, and 1000 bp larger, respectively. B. Chromosomal orc genes. Primers residing ~500 bp 5' and ~500 bp 3' to each chromosomally encoded orc gene (orc6, orc7, orc8, orc10) in Halobacterium sp. NRC-1 were used with total genomic DNA from individual colony isolates in PCR reactions to screen for orc gene knockouts. For orc6, orc7, orc8, and orc10, knockout alleles have a size of ~1000 bp where obtained, wild-type alleles are approximately 1100, 1500, 1200, and 1400 bp larger, respectively.

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