Paris/October 16

From 2007.igem.org

yesterday -- tomorrow

PCR rxn

the following PCR reactions have been performed

PCR : PrefixBB-frt1-CmR-frt2-SufixBB
PCR Settings Buffer (5x) 5x 10µL (GC rich) Expected size
Annealing (°C) MgCl2 10µM 10µM 0µL (no extra) 1Kb
0min dNTP 10µM 10µM 1µL Success
Time Elongation Oligo F 10µM O51 10µM 4µL YES
1m40' Oligo R 10µM O52 10µM 4µL Image (click to enlarge)
Number cycles Water qsp 50µL [[Image:|30px]]
30 Polymerase Phusion 0,5µL Band (0=ladder)
DNA pY-2P-IntC 5µL
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)
0min dNTP 10µM 10µM 1µL Success
Time Elongation Oligo F 10µM O51 10µM 4µL NO
1m40' Oligo R 10µM O53 10µM 4µL Image (click to enlarge)
Number cycles Water qsp 50µL [[Image:|30px]]
30 Polymerase Phusion 0,5µL Band (0=ladder)
DNA pY-2P-IntC 1µL (dil. 1/10)
PCR : PrefixBB-mutDGAT-SufixBB
PCR Settings Buffer (5x) 5x 10µL (GC rich) Expected size
Annealing (°C) MgCl2 10µM 10µM 0µL (no extra)
0min dNTP 10µM 10µM 1µL Success
Time Elongation Oligo F 10µM O29 10µM 4µL NO
2m Oligo R 10µM O30 10µM 4µL Image (click to enlarge)
Number cycles Water qsp 50µL [[Image:|30px]]
33 Polymerase Phusion 0,5µL Band (0=ladder)
DNA P13 1µL (dil. 1/10)