Purification method | Number of coloniesf
| Cloning efficiency |
---|
Prx IIE | G6PDH1 | Prx IIE | G6PDH1 |
---|
Nonea
| 86.3 ± 2.3 | 76.0 ± 4.0 | 15/18 | 17/18 |
EtOH pptb
| 546 ± 102 | 514 ± 68.7 | 18/18 | 17/18 |
ExoSAP-ITc
| 481 ± 117 | 507 ± 105 | 18/18 | 18/18 |
Columnd
| 7,170 ± 806 | 4,660 ± 1,400 | 18/18 | 18/18 |
Agarose gele
| 11,900 ± 1,750 | 2,830 ± 100 | 18/18 | 18/18 |
- The insert DNA fragments were amplified using 19-bp overlap primers and KOD DNA polymerase [51] (for Prx IIE gene) and PrimeSTAR Max DNA polymerase (for G6PDH1). The linearized vector DNA was prepared by PCR. The PCR solutions were treated with DpnI. The SLiCE (JM109) reaction was performed for 10 min at 37 °C. aUnpurified insert DNA was directly used for the SLiCE reaction. bInsert DNA was precipitated by ethanol. cPCR solution was treated with ExoSAP-IT (Affymetrix). dInsert DNA was purified from the PCR solution using a Gel/PCR Extraction Kit (FastGene). eInsert DNA was purified by agarose gel electrophoresis and using a Gel/PCR Extraction Kit (FastGene). An equivalent volume to 1/20 of the PCR solution was used for the SLiCE reaction. fEach value of “number of colonies” is the mean ± standard deviation of three independent experiments