IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Gold Bioremediation/2010/07/15

From OpenWetWare

Jump to: navigation, search
iGEM Project name 1 Main project page
Previous entry      Next entry

July 15th 2010

Digestion of pSB1A2 backbone, GolB, and GolS

GolB+GolS
30 ul template 2 ul Buffer 1 ul XbaI 1 ul SpeI 16 ul dH2O

pSB1A2
1.94 ul template 2 ul Buffer 1 ul XbaI 1 ul SpeI 44.06 ul dH2O

note: Should have included BSA and added 5 ul Buffer for 50 ul reaction

Gel Extraction of pSB1A2
No bands for GolS or GolB
Gel extracted backbone using Zymo kit, got good concentrations.

PCR-amplified GolB and GolS
41.1 ul H2O
0.5 ul template
1 ul Primer 1
1 ul Primer 2
1 ul dNTP
5 ul Buffer
0.4 ul polymerase

1. 95 degC for 5 min
2. 95 degC for 30 sec
3. 51 degC for 30 sec
4. 72 degC for 1 min
5. Repeat from step 2 for 31 cycles
6. 72 degC for 10 min




Personal tools