Table 4.

Summary of LR Reactions of Two- and Three-Segment Cloning

LR reaction (10 μL)	DNA topology	Input DNA (ng)	DNA size (bp)	fmoles	amp^R cfu 5 h^a	amp^R cfu 20 h^a
L1.1-STE2a-L3	linear	50	2768	27
R3-STE2bc-L2.1	linear	50	3713	20	71	238 (10/10)^b
pDEST425-B4-STE2 promoter	linear	150	9844	23
R1.1-ccdB-R2.1
L1.1-STE2a-R3	linear	50	2768	27
L3-STE2b-L4	linear	50	3340	22	149	330 (8/10)
R4-STE2c-L2.1	linear	50	2930	26
pDEST425-B4-STE2 promoter	linear	150	9844	23
R1.1-ccdB-R2.1
No Clonase						3

↵a Colony forming units per milliliter transformation obtained from transformation of 1 μL reaction into 50 μL DH5α competent cells at 1 × 10⁸ cfu/μg pUC.
↵b Number of correct clones vs. total number of clones analyzed based on restriction enzyme analysis of miniprep DNA.

Concerted Assembly and Cloning of Multiple DNA Segments Using In Vitro Site-Specific Recombination: Functional Analysis of Multi-Segment Expression Clones