From 2011.igem.org
SiL & GFP PCT
Project: Surface Display
Date: Tue, 30 Aug 2011 18:38:12 GMT
Author: Viki Ding (dingx055)
Access: Public
Revision History:
- Mon, 26 Sep 2011 18:47:02 GMT (dingx055): entry created in project'Surface Display' by dingx055 (id=9)
New vent polymerase ordered.
Objective: obtain SiL and GFP fragments
Silicatein (~1 kb)
- 1 uL vent pol.
- 3 uL dNTP
- 5 uL vent buffer
- 5 uL forward primer
- 5 uL reverse primer
- 2 uL SiL DNA
- 29 uL H2O
Taq mix
- 5 uL forward primer
- 5 uL reverse primer
- 2 uL SiL DNA
- 12 uL taq green mix
PCR conditions
95 degrees >> 4 min
95 degrees >> 30 sec
63 degrees >> 30 sec
72 degrees >> 1 min
34 cycles
72 degrees >> 10 min
GFP (less than 1 kb, ~950 bp)
- 1 uL vent pol.
- 3 uL dNTP
- 5 uL vent buffer
- 5 uL forward primer
- 5 uL reverse primer
- 2 uL GFP DNA
- 29 uL H2O
Taq mix
- 5 uL forward primer
- 5 uL reverse primer
- 2 uL GFP DNA
- 12 uL taq green mix
PCR conditions
95 degrees >> 4 min
95 degrees >> 30 sec
60 degrees >> 30 sec
72 degrees >> 1 min
34 cycles
72 degrees >> 10 min
Results
- Sil taq: thick bright band below 500 bp
- Sil vent: thick bright band at ~ 1 kb
- GFP taq: thick bright band at ~ 950 bp
- GFP vent: think bright band at ~ 950 bp
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