IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/07/14

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  • Made LB agar plates

1 L of distilled water, 35 grams of LB agar powder. Autoclave for 30 minutes.

  • Ran Digestion of TesA and RBS

TesA cut at X, P and RBS cut at S, P. Note: in order to ensure the viability of our PstI enzyme, we ran a control with some PstI from Haynes lab. Results are shown below. the PstI of Haynes lab had full digestion, while the iGEM PstI had a small amount of partial digestion, as made manifest by the three bands of the TesA in lane 2 of the gel.

Reagent RBS (μL) TesA (μL) Lanes
Our PstI:
1. 1kb ladder
2. TesA (651 bp)
3. RBS (2083)
Haynes PstI:
4. TesA (651 bp)
5. RBS (2083 bp)
6. Repeat of 5
DNA(plasmid) 10.0 10.0
10X FD buffer 2.0 2.0
Enzyme # 1 SpeI: 1.0 XbaI: 1.0
PstI 1.0 1.0
dH2O 6.0 6.0
  20 μL @ 42°C/ ~1 Hr.