Table 3. Efficiency of CDS cloning into pKK-RNAi vectors.
| Constructs attempted | First attempt at CDS assembly successful (fraction) |
First attempt at SLIC reaction successful (fraction) |
Correct sequence on n-th clone checked (fraction) |
|||||
|---|---|---|---|---|---|---|---|---|
| n = 1 | n = 2 | n = 3 | n = 4 | n = 5 | n = 6 | |||
| 45 | 40 (89%) |
42 (93%) |
34 (75.5%) |
5 (11%) |
2 (4.5%) |
2 (4.5%) |
1 (2.25%) |
1 (2.25%) |
Splice-PCR was used for in vitro assembly of miRNA-insensitive coding sequences that were subsequently cloned into the pKK-RNAi vector by our universal SLIC protocol.