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Table 1 Transformation Results

From: Amplified insert assembly: an optimized approach to standard assembly of BioBrickTM genetic circuits

Amplified Insert Assembly

#

Ligation Inputs

Results

Ā 

Insert

Vector

Ratio

Color

Correct

Incorrect

Background

% Accuracy

Ā 

Ptet

lacZ Ī±

1:1

Blue

819

6

1

99.6 Ā± 0.9%

1

Ptet

lacZ Ī±

2:1

Blue

868

13

1

98.4 Ā± 0.7%

Ā 

Ptet

lacZ Ī±

4:1

Blue

793

0

1

99.9 Ā± 0.2%

Ā 

PJ23100

lacZ Ī±

1:1

Blue

778

3

3

99.2 Ā± 0.6%

2

PJ23100

lacZ Ī±

2:1

Blue

1454

5

1

99.6 Ā± 0.8%

Ā 

PJ23100

lacZ Ī±

4:1

Blue

607

0

1

99.8 Ā± 0.3%

Ā 

lacZ Ī±

PJ23100

1:1

Blue

391

12

6

95.6 Ā± 2.2%

3

lacZ Ī±

PJ23100

2:1

Blue

547

2

3

99.1 Ā± 0.7%

Ā 

lacZ Ī±

PJ23100

4:1

Blue

465

5

1

98.7 Ā± 3.3%

Ā 

mRFP Driver

lacZ Ī±

1:1

Red

676

34

5

94.6 Ā± 3.4%

4

mRFP Driver

lacZ Ī±

2:1

Red

1329

24

1

98.2 Ā± 0.9%

Ā 

mRFP Driver

lacZ Ī±

4:1

Red

1761

22

6

98.4 Ā± 1.0%

Three-Antibiotic Assembly

#

Ligation Inputs

Results

Ā 

Prefix

Suffix

Vector

Color

Correct

Incorrect

% Accuracy

5

PJ23100

lacZ Ī±

pSB1C3

Blue

8

1

89%

5*

PJ23100

lacZ Ī±

pSB1C3

Blue

418

43

90.7 Ā± 0.6%

6

lacZ Ī±

mRFP Driver

pSB1C3

Red

3

1

75%

6*

lacZ Ī±

mRFP Driver

pSB1C3

Red

174

29

85.7 Ā± 4.6%

  1. The cumulative number of colonies after performing assemblies in triplicate on three separate days are shown. 3A ligations marked with an asterisk (*) were purified and concentrated before transformation. The highest % accuracy is bolded for each ligation reaction.