Table 2.
Cloning efficiency: numbers of colonies and of clones with correct length
| Viral Gene | Length (nt) | PCR amplicon (ng) | Transformed (µl) | Bacteria colonies | Correct clones/ Number analyzed |
|---|---|---|---|---|---|
| PB2 | 2341 | 120 | 1 | 0 | 11/12 |
| 120 | 10 | 34 | |||
| PB1 | 2341 | 40 | 1 | 60 | 14/16 |
| 40 | 10 | 20 | |||
| PA | 2233 | 600 | 1 | 5 | 6/8 |
| 600 | 10 | 92 | |||
| HA | 1779 | 490 | 1 | 26 | 8/8 |
| 490 | 10 | 65 | |||
| NP | 1565 | 500 | 1 | 23 | 8/8 |
| 500 | 10 | 48 | |||
| NA | 1399 | 430 | 1 | 18 | 4/5 |
| 430 | 10 | 39 | |||
| M | 1027 | 150 | 1 | 12 | 8/8 |
| 150 | 10 | 140 | |||
| NS | 875 | 690 | 1 | 19 | 12/12 |
| 690 | 10 | 150 |
Numbers of bacteria colonies (XL1-Blue™ bacteria, in case of the PB1 gene SURE2™ bacteria) and of positive plasmid clones carrying the cDNA of viral genes from strain A/Thailand/1(KAN-1)/2004 (H5N1) with correct length in relation to viral gene length, approximate amounts of PCR amplicons used for target-primed plasmid amplification and amounts of it used for transformation.