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Table 1 Estimation of the crossover number for β-lactamase. The number of colonies with reactivated β-lactamase (functional) is given as a percentage.

From: Revealing biases inherent in recombination protocols

  

RD-PCR

A/E temp =

 

DNA-shuffling

  

40*

45*

60+

60+

60+

  

A/E time

CR/1000 bp

5 sec

5 sec

5 sec

15 sec

5 sec-Pfu

CR/1000 bp

< 150 bp

Min-1

1.3

59.2% ± 6.1

63.4% ± 6.6

73.3% ± 7.1

59.8% ± 6.4

66.9% ± 4.4

1.3

61.7% ± 4.3

Min-2/3

2.6

30.0% ± 4.9

33.8% ± 3.5

59.7% ± 3.8

45.6% ± 4.2

46.5% ± 2.7

2.7

38.1% ± 2.7

Min-3

4.1

19.6% ± 3.6

22.5% ± 2.9

44.0% ± 4.3

28.4% ± 3.1

35.4% ± 3.2

4.1

25.3% ± 3.7

Min-4/5

5.3

9.9% ± 2.5

13.7% ± 2.2

29.8% ± 3.3

17.6% ± 2.5

27.7% ± 2.7

5.4

16.6% ± 2.9

  1. CR = crossovers required to gain activity
  2. A/E = annealing/extension
  3. *94°C for 5 min; 40 cycles of (94°C, 30 s; 40–45°C, 5 s; 72°C for 3 s); 10 cycles of (94°C, 30 s; 50°C, 30 s; and 72°C 30 s); + 94°C 2 min, 99 cycles of (94°C 1 min, 63–67°C, 5 sec), 72°C 7 min, hold at 4°C
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BMC Biotechnology

ISSN: 1472-6750

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