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From 2011.igem.org

Contents 1 23. Labday 2 Digestion of PCR products 3 Sequencing 4 3A-assembly: Digestion of plastic-binding site and GFP

23. Labday

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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.

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.

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

Investigators: Sophie, Sandra

1. Digestion of GFP-plastic-binding site part

• Enzyme 1: XbaI
• Enzyme 2: SpeI

1. Digestion of Promotor-RBS part

• EcoRI
• SpeI

1. Digestion of Vector pSB1K3

• Enzyme 1: EcoRI
• Enzyme 2: SpeI