Paris/October 20
From 2007.igem.org
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== PCR reaction == | == PCR reaction == | ||
| + | |||
| + | {{Paris_PCR_0| Title = PrefixBB-frt1-CmR-frt2-T1T2-SufixBB | ||
| + | |Annealing= 60 | ||
| + | |Elongation= 2'30 | ||
| + | |Cycles= 33 | ||
| + | |Buffer= 5x 10µL (GC rich) | ||
| + | |MgCl2= 10µM 0µL (no extra) | ||
| + | |dNTP= 10µM 1µL | ||
| + | |n_oligoF= O51 | ||
| + | |v_oligoF= 10µM 2.5µL | ||
| + | |n_oligoR= O53 | ||
| + | |v_oligoR= 10µM 2.5µL | ||
| + | |water= qsp 50µL | ||
| + | |pol= Phusion 0,5µL | ||
| + | |DNA= pY-2P-IntC 1µL (dil. 1/10) | ||
| + | |Size= | ||
| + | |Success=NO | ||
| + | |Image= | ||
| + | |Band= | ||
| + | |}} | ||
== Purification of Digestion and PCR products == | == Purification of Digestion and PCR products == | ||
Revision as of 13:05, 22 October 2007
Contents |
Digestion Reaction
PCR reaction
| PCR : PrefixBB-frt1-CmR-frt2-T1T2-SufixBB | ||||||
|---|---|---|---|---|---|---|
| PCR Settings | Buffer (5x) | 5x 10µL (GC rich) | Expected size | |||
| Annealing (°C) | MgCl2 10µM | 10µM 0µL (no extra) | ||||
| 60 | dNTP 10µM | 10µM 1µL | Success | |||
| Time Elongation | Oligo F 10µM | O51 | 10µM 2.5µL | NO | ||
| 2'30 | Oligo R 10µM | O53 | 10µM 2.5µL | Image (click to enlarge) | ||
| Number cycles | Water | qsp 50µL | [[Image:|30px]] | |||
| 33 | Polymerase | Phusion 0,5µL | Band (0=ladder) | |||
| DNA | pY-2P-IntC 1µL (dil. 1/10) | |||||
Purification of Digestion and PCR products
Ligation Reactions
Culture
Launching of 80mL culture of the strain countaining the plasmid pY-2P-IntC, last minipreps revealed the weak nomber of plasmid recover with a classic miniprep on 5mL culture.
