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: <br>
 +
Forward : Pref_ATG_Fw <br>
 +
Reverse : Prt D_Rv_Bal1 <br><br>
 +
 +
DNA : <br>
 +
Purified and amplified D protein from Lambda phage. <br><br>
 +
 +
Cycles: <br>
 +
94°C 2minutes; <br>
 +
(94°C 1 minute; <br>
 +
46°C 1 minutes; <br>
 +
72°C 1 minute(s)) X 35; <br>
 +
72°C 4 minutes; <br>
 +
 +
==== Second sample ====
 +
 +
Primers : <br>
 +
Forward : Pref_ATG_Fw <br>
 +
Reverse : Suffixe_Rv <br><br>
 +
 +
DNA : <br>
 +
Protéine D amplifiée et purifiée. <br><br>
 +
 +
Cycles: <br>
 +
94°C 2minutes; <br>
 +
(94°C 1 minute; <br>
 +
59°C 1 minutes; <br>
 +
72°C 1 minute(s)) X 35; <br>
 +
72°C 4 minutes; <br>
 +
 +
==== Penton base from adenovirus 5 ====
 +
 +
==== First sample ====
 +
 +
Primers: <br>
 +
Forward : Bal1_ADV5_Fw <br>
 +
Reverse : Bal1_ADV5_Rv <br><br>
 +
 +
DNA : <br>
 +
Purified and amplified penton base from adénovirus 5. <br><br>
 +
 +
Cycles: <br>
 +
94°C 2minutes; <br>
 +
(94°C 1 minute; <br>
 +
57,3°C 1 minutes; <br>
 +
72°C 2 minute(s)) X 35; <br>
 +
72°C 5 minutes; <br>
 +
 +
==== Second sample ====
 +
 +
Primers: <br>
 +
Forward : Pref_ATG_Fw <br>
 +
Reverse : Suffixe_Rv <br><br>
 +
 +
DNA : <br>
 +
Purified and amplified penton base from adénovirus 5. <br><br>
 +
 +
Cycles: <br>
 +
94°C 2minutes; <br>
 +
(94°C 1 minute; <br>
 +
59°C 1 minutes; <br>
 +
72°C 2 minute(s)) X 35; <br>
 +
72°C 5 minutes; <br>
 +
 +
==== DNA electrophoresis ====
 +
 +
85 Volt, 15 minutes. <br>
 +
105 Volt, 40 minutes. <br>
 +
Ladder fermentas 1 Kb. <br>
 +
 +
 +
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).
 +
<br>
 +
 +
[[image:M2109.png|center]]
 +
<br>
 +
 +
==== DNA purification ====
 +
 +
Kit Qiagen “gel extraction kit”, final volume = 50µL. <br>

Latest revision as of 15:06, 21 October 2009

framless



August
MTWTFSS
          [http://2009.igem.org/User:DavidC/1_August_2009 1] [http://2009.igem.org/User:DavidC/2_August_2009 2]
[http://2009.igem.org/User:DavidC/3_August_2009 3] [http://2009.igem.org/User:DavidC/4_August_2009 4] [http://2009.igem.org/User:DavidC/5_August_2009 5] [http://2009.igem.org/User:DavidC/6_August_2009 6] [http://2009.igem.org/User:DavidC/7_August_2009 7] [http://2009.igem.org/User:DavidC/8_August_2009 8] [http://2009.igem.org/User:DavidC/9_August_2009 9]
[http://2009.igem.org/User:DavidC/10_August_2009 10] [http://2009.igem.org/User:DavidC/11_August_2009 11] [http://2009.igem.org/User:DavidC/12_August_2009 12] [http://2009.igem.org/User:DavidC/13_August_2009 13] [http://2009.igem.org/User:DavidC/14_August_2009 14] [http://2009.igem.org/User:DavidC/15_August_2009 15] [http://2009.igem.org/User:DavidC/16_August_2009 16]
[http://2009.igem.org/User:DavidC/17_August_2009 17] [http://2009.igem.org/User:DavidC/18_August_2009 18] [http://2009.igem.org/User:DavidC/19_August_2009 19] [http://2009.igem.org/User:DavidC/20_August_2009 20] [http://2009.igem.org/User:DavidC/21_August_2009 21] [http://2009.igem.org/User:DavidC/22_August_2009 22] [http://2009.igem.org/User:DavidC/23_August_2009 23]
[http://2009.igem.org/User:DavidC/24_August_2009 24] [http://2009.igem.org/User:DavidC/25_August_2009 25] [http://2009.igem.org/User:DavidC/26_August_2009 26] [http://2009.igem.org/User:DavidC/27_August_2009 27] [http://2009.igem.org/User:DavidC/28_August_2009 28] [http://2009.igem.org/User:DavidC/29_August_2009 29] [http://2009.igem.org/User:DavidC/30_August_2009 30]
[http://2009.igem.org/User:DavidC/31_August_2009 31]
September
MTWTFSS
  [http://2009.igem.org/User:DavidC/1_September_2009 1] [http://2009.igem.org/User:DavidC/2_September_2009 2] [http://2009.igem.org/User:DavidC/3_September_2009 3] [http://2009.igem.org/User:DavidC/4_September_2009 4] [http://2009.igem.org/User:DavidC/5_September_2009 5] [http://2009.igem.org/User:DavidC/6_September_2009 6]
[http://2009.igem.org/User:DavidC/7_September_2009 7] [http://2009.igem.org/User:DavidC/8_September_2009 8] [http://2009.igem.org/User:DavidC/9_September_2009 9] [http://2009.igem.org/User:DavidC/10_September_2009 10] [http://2009.igem.org/User:DavidC/11_September_2009 11] [http://2009.igem.org/User:DavidC/12_September_2009 12] [http://2009.igem.org/User:DavidC/13_September_2009 13]
[http://2009.igem.org/User:DavidC/14_September_2009 14] [http://2009.igem.org/User:DavidC/15_September_2009 15] [http://2009.igem.org/User:DavidC/16_September_2009 16] [http://2009.igem.org/User:DavidC/17_September_2009 17] [http://2009.igem.org/User:DavidC/18_September_2009 18] [http://2009.igem.org/User:DavidC/19_September_2009 19] [http://2009.igem.org/User:DavidC/20_September_2009 20]
[http://2009.igem.org/User:DavidC/21_September_2009 21] [http://2009.igem.org/User:DavidC/22_September_2009 22] [http://2009.igem.org/User:DavidC/23_September_2009 23] [http://2009.igem.org/User:DavidC/24_September_2009 24] [http://2009.igem.org/User:DavidC/25_September_2009 25] [http://2009.igem.org/User:DavidC/26_September_2009 26] [http://2009.igem.org/User:DavidC/27_September_2009 27]
[http://2009.igem.org/User:DavidC/28_September_2009 28] [http://2009.igem.org/User:DavidC/29_September_2009 29] [http://2009.igem.org/User:DavidC/30_September_2009 30]
October
MTWTFSS
      [http://2009.igem.org/User:DavidC/1_October_2009 1] [http://2009.igem.org/User:DavidC/2_October_2009 2] [http://2009.igem.org/User:DavidC/3_October_2009 3] [http://2009.igem.org/User:DavidC/4_October_2009 4]
[http://2009.igem.org/User:DavidC/5_October_2009 5] [http://2009.igem.org/User:DavidC/6_October_2009 6] [http://2009.igem.org/User:DavidC/7_October_2009 7] [http://2009.igem.org/User:DavidC/8_October_2009 8] [http://2009.igem.org/User:DavidC/9_October_2009 9] [http://2009.igem.org/User:DavidC/10_October_2009 10] [http://2009.igem.org/User:DavidC/11_October_2009 11]
[http://2009.igem.org/User:DavidC/12_October_2009 12] [http://2009.igem.org/User:DavidC/13_October_2009 13] [http://2009.igem.org/User:DavidC/14_October_2009 14] [http://2009.igem.org/User:DavidC/15_October_2009 15] [http://2009.igem.org/User:DavidC/16_October_2009 16] [http://2009.igem.org/User:DavidC/17_October_2009 17] [http://2009.igem.org/User:DavidC/18_October_2009 18]
[http://2009.igem.org/User:DavidC/19_October_2009 19] [http://2009.igem.org/User:DavidC/20_October_2009 20] [http://2009.igem.org/User:DavidC/21_October_2009 21] [http://2009.igem.org/User:DavidC/22_October_2009 22] [http://2009.igem.org/User:DavidC/23_October_2009 23] [http://2009.igem.org/User:DavidC/24_October_2009 24] [http://2009.igem.org/User:DavidC/25_October_2009 25]
[http://2009.igem.org/User:DavidC/26_October_2009 26] [http://2009.igem.org/User:DavidC/27_October_2009 27] [http://2009.igem.org/User:DavidC/28_October_2009 28] [http://2009.igem.org/User:DavidC/29_October_2009 29] [http://2009.igem.org/User:DavidC/30_October_2009 30] [http://2009.igem.org/User:DavidC/31_October_2009 31]


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.

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