User:DavidC/21 September 2009
From 2009.igem.org
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Monday the 21th
PCR
Fusion protein design
D protein
First sample
Primers:
Forward : Pref_ATG_Fw
Reverse : Prt D_Rv_Bal1
DNA :
Purified and amplified D protein from Lambda phage.
Cycles:
94°C 2minutes;
(94°C 1 minute;
46°C 1 minutes;
72°C 1 minute(s)) X 35;
72°C 4 minutes;
Second sample
Primers :
Forward : Pref_ATG_Fw
Reverse : Suffixe_Rv
DNA :
Protéine D amplifiée et purifiée.
Cycles:
94°C 2minutes;
(94°C 1 minute;
59°C 1 minutes;
72°C 1 minute(s)) X 35;
72°C 4 minutes;
Penton base from adenovirus 5
First sample
Primers:
Forward : Bal1_ADV5_Fw
Reverse : Bal1_ADV5_Rv
DNA :
Purified and amplified penton base from adénovirus 5.
Cycles:
94°C 2minutes;
(94°C 1 minute;
57,3°C 1 minutes;
72°C 2 minute(s)) X 35;
72°C 5 minutes;
Second sample
Primers:
Forward : Pref_ATG_Fw
Reverse : Suffixe_Rv
DNA :
Purified and amplified penton base from adénovirus 5.
Cycles:
94°C 2minutes;
(94°C 1 minute;
59°C 1 minutes;
72°C 2 minute(s)) X 35;
72°C 5 minutes;
DNA electrophoresis
85 Volt, 15 minutes.
105 Volt, 40 minutes.
Ladder fermentas 1 Kb.
Samples: D protein (D prt BBa) with BioBrick prefix and suffix, adenovirus penton base with BioBrick prefix and suffix (ADV5pb BBa), D protein with Biobriock prefix and BalI sequence into the suffix (D prt BalI), adenovirus penton base with Biobrick suffix and BalI sequence into the prefix (ADV5pb BalI).
DNA purification
Kit Qiagen “gel extraction kit”, final volume = 50µL.