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From 2011.igem.org

Contents 1 green light receptor 1.1 Digestion of PCR products 2 Lysis cassette 2.1 Sequencing 3 Precipitator 3.1 3A-assembly: Digestion of plastic-binding site and GFP

green light receptor

Digestion of PCR products

Investigators: Julia

Digest of PCR product Ccas.

Ccas:

• Enzyme 1: XbaI
• Enzyme 2: PstI

in vector pSB1C3:

• Enzyme 1: SpeI
• Enzyme 2: PstI

Comments: RFP is not digested with SpeI and PstI. It was digested again, that time with NEB4 and XbaI.

Lysis cassette

Sequencing

Investigators: Theo

PCR of Lys1, Lys2, Lys3, Lys4, Lys5.

Materials:

• H2O
• Seq. Buffer
• DNA (Lys 1-Lys5)
• Primer
• Polymerase

PCR programm:

1. 95°C 3 min
2. 95°C 15 sec
3. 50°C 15 sec
4. 60°C 2 min
5. 4°C stored til needed again

step 2 to 4 was repeated 40x times.

Precipitator

3A-assembly: Digestion of plastic-binding site and GFP

Investigators: Sophie, Sandra

1. Digestion of GFP-plastic-binding site part (PCR product)
   • Enzyme 1: XbaI
   • Enzyme 2: SpeI

1. Digestion of Promotor-RBS part (PR1-PR6)
   • EcoRI
   • SpeI

1. Digestion of Vector pSB1K3
   • Enzyme 1: EcoRI
   • Enzyme 2: SpeI