IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Output/2009/07/03/Zhangs

From OpenWetWare

Jump to: navigation, search

Add CIAP into 12 RBS digesting vector samples.
Identify the digested ligation product of CI+1-23L/1-4H and RBS vector by electrophoresis
System:

1μLsample
1μLDNA Dye
4μLddH2O

Marker:

5μLMarker
1μLDNA Dye

Digestions for RBS vectors failed and we digested again.
Samples: 1-11N, 1-1H, 1004-4C, 1-2M, 1-2I, 1-5N, 1-2G, 1-2K, 1-5J, 1004-3C, 1004-2G, 1-1J
System

6μLRBS plasmid
1.5μLSpeI
1.5μLPstI
2μL10×H buffer
9μLddH2O
10μLTotal

Gel Extraction for CI+1-23L/1-4H insert
Gel: CI+1-23L 0.139gPN volume: 417μL
CI+1-4H 0.071gPN volume:213μL

Identify the RBS digestion products by electrophoresis
Results (from left to right)
Plasmid control, 1-2I, 1004-2G, 1004-4C, 1-5N, 1004-3C, 1-2K, 1-1H, 1-1J, 1-2G, 1-11N, 1-2M, 1-5J

There are no strips for thee 2008 iGEM parts 1004-2G, 1004-4C, 1004-3C. Perhaps the quality of the 2008 parts is bad.

DNA purification to extract RBS vector

Ligate the RBS vector and CI+Terminator insert
System

3μLinsert
1μLvector
1μL10× Ligase buffer
1μLLigase
4μLddH2O
10μLTotal
Personal tools