Chi is a hotspot for homologous recombination mediated by the RecBCD (Rec) pathway of Escherichia coli. For Rec-mediated recombination of phage λ, the orientation of λ cos in the λ chromosome dictates the direction of travel of RecBCD enzyme through DNA and dictates which orientation of Chi or Chi-like sequences will be active in stimulating recombination. I previously found that Rec-mediated λ patch heteroduplexes, stimulated by Chi or not, are chain-biased; at the λ P locus, recombinant information resides on the λ r chain. This bias exists in the presence or absence of Chi sites. Reported herein is the finding that r-chain-bias at the P locus is independent of the orientation of λ cos and thus also independent of the orientation of active Chi's or Chi-like sequences and of the direction of travel of RecBCD enzyme. These results disprove previously elaborated models in which a chain-specific nick at Chi initiates recombination, and imply that some other chain-distinguishing process is involved with recombination. Replication and transcription are candidates for such a process.