Table 1

Testing of USER Friendly cloning for single step four fragment cloning

No. of correct colonies/total no. of colonies

Average

Vector

Insert 1

Insert 2

1.

2.

3.

pRF-HU

PKS1-A1/A2

na

87/91

100/103

114/120

104.7 (95.9%)

pRF-HUE

PKS1-O1/O2

na

59/61

60/63

66/70

64.7 (95.4%)

pRF-HU2

PKS1-A1/A2

PKS1-A3/A4

47/53

52/61

45/55

56.3 (85.2%)

pRF-HU2E

PKS1-O1/O2

PKS1-O3/O4

38/48

39/45

40/49

47.3 (82.4%)

USER cloning: Results from three independent replicates where one or two PCR amplicons are cloned into the designed USER vectors. All transformants were first tested for the presence of the desired insert and then for the orientation of the insert in the vector. All transformants that were positive for inserts also had the correct orientation. The negative control reactions, where no PCR amplicons were added to the reactions, resulted in zero colonies.

Frandsen et al. BMC Molecular Biology 2008 9:70   doi:10.1186/1471-2199-9-70