User:DavidC/21 September 2009

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framless



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Contents

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).

M2109.png


DNA purification

Kit Qiagen “gel extraction kit”, final volume = 50µL.