"
Team:Freiburg/Notebook/4 July
From 2011.igem.org
(Difference between revisions)
(→Digestion of PCR products) |
(→Sequencing) |
||
| Line 15: | Line 15: | ||
<span style="color:blue;">Comments: RFP is not digested with SpeI and PstI. It was digested again, that time with NEB4 and XbaI.</Span> | <span style="color:blue;">Comments: RFP is not digested with SpeI and PstI. It was digested again, that time with NEB4 and XbaI.</Span> | ||
| - | ==<span style="color:orange;"> | + | ==<span style="color:orange;">Lysis cassette</span>== |
| - | + | ===Sequencing=== | |
'''Investigators: Theo''' | '''Investigators: Theo''' | ||
| Line 36: | Line 36: | ||
step 2 to 4 was repeated 40x times. | step 2 to 4 was repeated 40x times. | ||
| - | |||
==<span style="color:grey;">3A-assembly: Digestion of plastic-binding site and GFP</span>== | ==<span style="color:grey;">3A-assembly: Digestion of plastic-binding site and GFP</span>== | ||
Revision as of 10:19, 15 July 2011
Contents |
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:
- 95°C 3 min
- 95°C 15 sec
- 50°C 15 sec
- 60°C 2 min
- 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
- Digestion of GFP-plastic-binding site part (PCR product)
- Enzyme 1: XbaI
- Enzyme 2: SpeI
- Digestion of Promotor-RBS part (PR1-PR6)
- EcoRI
- SpeI
- Digestion of Vector pSB1K3
- Enzyme 1: EcoRI
- Enzyme 2: SpeI
