IGEM:UNAM-Genomics Mexico/2009/Notebook/H2 Pae/2011/08/22

iGEM Project name 1

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Abstract Continuing the assembly of the parts which are going to be sent to the registry. Parts assembly Today I did a PCR from the pSB1C3 plasmid amplifying all the backbone, because I will need it to isert our parts. I did the PCR with the RTTH enzyme, according to the following steps. 1. I prepared the two mixes, which were: Mix 1 Compound Volume(μL) H20 9 Buffer rTth[3.3x] 6 Mg(OAc)2[25mM] 3 dNTPs[10mM] 3 Oligo UP[5 pmol/μL] 4 Oligo DW[5 pmol/μL] 4 Diluted DNA (1/50) 1 Mix 2 Compound Volume(μL) H20 10.5 Buffer rTth 9 rTth DNApol 0.5 2. Once ready a I put the mix into the thermocylcer, the first cycle was: 94°C/5min 3. Then I added the mix 2, after that the following cycle were done: 30 cylces: 94°C / 30 seg 60°C / 30 seg 72°C / 3 min 1 cycle: 72°C / 5 min 4° C / ∞ I will see the results on wednesday, in an electrophoresis gel.