Summary of LR Reactions of Two- and Three-Segment Cloning
LR reaction (10 μL) | DNA topology | Input DNA (ng) | DNA size (bp) | fmoles | ampR cfu 5 ha | ampR cfu 20 ha |
|---|---|---|---|---|---|---|
| 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.