Table 1.
Screening Efficiency of Gateway Cloning
|
|
Panel of selected ORFs |
||
|---|---|---|---|
|
|
A
|
B
|
|
| ORF size range (bp) | 500-3000 | 200-5500 | |
| Number of ORFs screened | 111 | 192 | |
| 4/4 positive colonies | 72 | 108 | |
| 3/4 positive colonies | 11 | 28 | |
| 2/4 positive colonies | 06 | 15 | |
| 1/4 positive colonies | 06 | 08 | |
| 0/4 positive colonies | 16 | 33 | |
| Cloning efficiency (%)
|
86
|
83
|
|
-
The screening for recombinant master clones was performed by colony PCR on 4 colonies per each BP (gene) reaction. A positive colony represented the correct amplified gene size analyzed on agarose gel. DNA sequencing using plasmid-specific primers later confirmed the identity of the cloned genes.
