Colony PCR

Checked if primers PS-R, EX-F were good to use for colony PCR and also checked if insert was realy in plasmids

• September 6th transformation (digestion was done with H buffer×２ and M buffer×２, 2 version)
• 1-3A
• Retried September 6th transformation
• pUC119

Confirmation of pSB1C3

Because pSB1C3 PCRed from 1-3A（RFP reporter） was used there was posibility of contamination by template which would also produce red colonies.

• H buffer treated 2 uL ＋ Mighty Mix 2 uL ＋ T4 ligase 0.5 uL　→　Ligation　→　Transformation
• M buffer treated 2 uL ＋ Mighty Mix 2 uL ＋ T4 ligase 0.5 uL　→　Ligation　→　Transformation
• H buffer treated 2 uL　→　Transformation
• M buffer treated 2 uL　→　Transformation

PCR of GFP

• GFP reporter BBa_I13522 pSB1A2 2-8A 937 bp
• GFP protein BBa_E0840 pSB1A2 1-12O 878 bp

PCR cocktail was same as September 1st's