Cleavage and polyadenylation specificity factor 30 (CPSF30) is a 'zinc finger' protein that plays a crucial role in the transition of pre-mRNA to RNA. CPSF30 contains five conserved CCCH domains and a CCHC "Zinc Knuckle" domain. CPSF30 activity is critical for pre-mRNA processing. A truncated form of the protein, in which only the CCCH domains are present, has been shown to specifically bind AU-rich pre-mRNA targets; however, the RNA binding and recognition properties of full length CPSF30 are not known. Herein, we report the isolation and biochemical characterization of full length CPSF30. We report that CPSF30 contains one 2Fe-2S cluster in addition to five zinc ions, as measured by ICP-MS, UV-visible and XAS spectroscopies. Utilizing fluorescence anisotropy RNA binding assays, we show that full length CPSF30 has high binding affinity for two types of pre-mRNA targets -AAUAAA and poly U -both of which are conserved sequence motifs present in the majority of pre-mRNAs. Binding to the AAUAAA motif requires that the 5 CCCH domains of CPSF30 be present; whereas, binding to poly U sequences requires the entire, full length CPSF30. These findings implicate the CCHC "Zinc Knuckle" present in the full-length protein as critical for mediating poly U binding. We also report that truncated forms of the protein, containing either just two CCCH domains (ZF2 and ZF3) or the CCHC 'zinc knuckle' domain, do not exhibit any RNA binding, indicating that CPSF30/RNA binding requires several ZF (and/or Fe-S cluster) domains working in concert to mediate RNA recognition.