Objectives
- Gel extraccion of PCRs
- PCR for Insert. Again! Low concentration in gel extraction.
- Quiagen Gel Extraction Kit, with elution 30μL.
- Lane 1 Ladder.
- Lane 2 Insert with Prefix and suffix Standard for Gibson
- Lane 3 Backbone with Prefix and suffix complete.
I need to repeat the insert PCR and make a new Gel extraction for that biopart.
PCR for inserts
| Reagents
|
PCR 1 P1A7/2011
|
PCR 2 P1A7/2011
|
PCR 3 P1A7/2011(Control+)
|
| DNA
|
1μL
|
1μL
|
1μL
|
| Buffer 5x PhusionHF
|
10μL
|
10μL
|
10μL
|
| dNTPs(8mM)
|
5μL
|
5μL
|
5μL
|
| Oligo(Extnd_Pref_Fwd)
|
5μL
|
5μL
|
5μL‡
|
| Oligo(Extnd_Suff_Rev)
|
5μL
|
5μL
|
5μL‡
|
| Phusion
|
.5μL
|
.5μL
|
.5μL
|
| H2O
|
23.5μL
|
23.5μL
|
23.5μL
|
‡Prefix_Fwd and Suffix_Reverse Primers.
|
iGEM 2011
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