Team:NCKU Tainan/Protocol

Protocol

1. Gently mix the following reaction by pipetting and centrifuge briefly.

　 　 　 　 　 　 　 　 　 　 　 　 　 　 　
20 μl system 50 μl system
Template12～20 ng 30～50 ng
Forward primer1.0 μl 2.5 μl
Reverse primer1.0 μl 2.5 μl
dNTP1.6 μl 4.0 μl
10x Buffer2.0 μl 5.0 μl

Program of KOD DNA polymerase

　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　
TemperatureTime
94 ℃3 min. 25～30 cycles
94 ℃
(Denaturation)		40 sec
57.5 ℃
(Annealing)		30 sec
72 ℃
(Extension)		Depend on sequence size
(2 kbp/min. for Taq)
72 ℃5 min.
4 ℃∞

2. Confirm the size of the digested product by gel electrophoresis.
3. Gel purification of the target size.

Arduino

Arduino

Follow us
Contact us

igem.ncku.tainan@gmail.com
No.1, Daxue Rd., East Dist., Tainan City 701, Taiwan (R.O.C.)