Team:Uppsala-SwedenWeek8

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Week-8

Preparation for getting the bricks:

(1)Competent cell preparation

(2)Transformation of the Biobricks into DH5-alpha competent cells

(3)Verificiation of the lengths of the biobricks by Gel Electrophoresis after PCR.

Details of the biobricks we are using:

Biobricks details

Part BBa_

Name

Length (bp)

Plasmid Length

Total Length

Plasmid

Plasmid number

Plate

Well

Antibiotics

Sequencing

Quality (Gel, Q, P)

Function

F 2621

F2

1158

2079

3237

PSB1A2

All

2

21F

Amp

Confirmed

Bad, OK, OK

Sensor

F 1610

F1

798

3189

3987

pSB1Ak3

AHL Relay

2

24G

Amp & Km

Confirmed

OK, OK, OK

AHL generator

I 746350

F17

237

2079

2316

pSB1A2

1 & 4

2

12C

Amp

Partially

OK, OK, OK

Activator in sensitivity tuner

I 746352

F4

264

2079

2343

pSB1A2

2,3,5 & 6

2

12G

Amp

Partially

OK, OK, OK

Activator in sensitivity tuner

B0015

F5

129

3189

3318

pSB1AK3

All

1

23L

Amp & Km

Confirmed

OK, OK, OK

Terminator

I746360

F6

91

2079

2170

pSB1A2

1 & 4

2

12I

Amp

Confirmed

OK, OK, OK

Lower level promotor

I746364

F7

93

2079

2172

pSB1A2

2 & 5

2

12O

Amp

Confirmed

OK, OK, OK

Lower level promotor

I746365

F8

92

2079

2171

pSB1A2

3 & 6

2

14A

Amp

Inconsistent

OK, OK, OK

Lower level promotor

Q04510

F9

987

4425

5412

pSB2K3

All

1

18B

Km

Confirmed

OK, Low, OK

Inverter

P0412

F10

1308

2079

3387

pSB1A2

All

3

6P

Amp

Confirmed

OK, OK, OK

Lower level repressor

I746361

F11

92

2079

2171

pSB1A2

1,2,4 & 5

2

12K

Amp

Confirmed

OK, OK, OK

Higher level promotor

R0010

F12

200

2079

2279

pSB1A2

4,5 & 6

1

1D

Amp

Confirmed

OK, OK, OK

Regulatory promotor

I0460

F16

969

3189

4158

pSB1AK3

4,5 & 6

2

24I

Amp & Km

Partially

OK, OK, OK

AiiA generator

I13504

F13

875

2079

2954

pSB1A2

1 (GFP)

1

22I

Amp

Confirmed

OK, High, OK

GFP Generator

I13507

F14

861

2079

2940

pSB1A2

2 (RFP)

1

22O

Amp

Confirmed

OK, High, OK

RFP Generator

E0430

F15

878

2079

2957

pSB1A2

3 (YFP)

1

8I

Amp

Confirmed

OK, High, OK

YFP Generator

B0034

F3

12

2079

2091

pSB1A2

4,5 & 6

1

2M

Amp

Confirmed

OK, High, OK

RBS

Gel image of the biobricks after confirmation of lengths:

Biobricks length confirmation


* Outcome:We confirmed the lengths of almost all the transformed cells of the required biobricks to be correct as per above gel image.