Molecular analysis of gene transfer efficiency in male and female mice. (A) Southern blot analysis of undigested low-molecular-weight Hirt DNA extracted from liver 3 days (3 d), 10 days (10 d), and 20 days (20 d) after portal vein infusion of vector particles. C indicates DNA from naive control mouse; M and F, DNA from rAAV CAGG-FIX transduced male and female mice, respectively. (B) Southern blot analysis of high-molecular-weight genomic DNA isolated from the liver 10 weeks after portal vein infusion of 1 × 10¹¹ rAAV-2 CAGG-FIX vector and digested with EcoRI and HindIII to release a 3.3-kb fragment from the expression cassette. C indicates DNA from naive control mouse; M and F, DNA from rAAV CAGG-FIX transduced male and female mice, respectively. The final 2 right-hand lanes consist of control mouse DNA spiked with 4.2 and 0.42 copies per diploid genome of plasmid pAV CAGG-FIX digested with EcoRI and Hind III. (C) Northern blot analysis of total cellular RNA isolated from liver of male (M) and female (F) mice, 10 weeks following portal vein infusion of 1 × 10¹¹ rAAV CAGG-FIX vector. C indicates RNA from naive control male mouse. For each of the above procedures, equivalent loading of nucleic acid on the respective gels was documented using AlphaImager software version 3.24 (Alpha Innotech).