Team:Heidelberg/Notebook/ViroBytes/October
From 2010.igem.org
(→27/09/2010) |
(→15/10/2010) |
||
Line 87: | Line 87: | ||
==15/10/2010== | ==15/10/2010== | ||
- | Fragments 1-4; 5-8 assembly. | + | **Fragments 1-4; 5-8 assembly. |
- | [[Image:Gel_546.jpg|thumb| | + | [[Image:Gel_546.jpg|thumb|400px|right|]] |
*1 - fragments 1-4 assembly. | *1 - fragments 1-4 assembly. | ||
Line 96: | Line 96: | ||
*3 - assembly ligation mix | *3 - assembly ligation mix | ||
M - DNA 1kb ladder | M - DNA 1kb ladder | ||
- | *4 - fragments | + | *4 - fragments 5-8 assembly. |
*5 - fragments control. | *5 - fragments control. | ||
*6 - assembly ligation mix | *6 - assembly ligation mix | ||
+ | |||
+ | ==18/10/2010== | ||
+ | |||
+ | **Fragments 1-4; | ||
+ | |||
+ | |||
+ | [[Image:Gel_547.png|thumb|500px|right|]] | ||
+ | |||
+ | M - DNA 1kb ladder | ||
+ | *1 - control (fragments 1-4 from AAV1). | ||
+ | *2 - fragments 1-4 assembly | ||
+ | *3 - fragments 1-4 assembly (2nd reaction) | ||
+ | *4 - negative control | ||
==25/10/2010== | ==25/10/2010== |
Revision as of 03:39, 28 October 2010
![](https://static.igem.org/mediawiki/2010/9/92/Bg_plasmid.png)
October4/10/2010
5/10/2010
15/10/2010
M - DNA 1kb ladder
18/10/2010
M - DNA 1kb ladder
25/10/2010
PCR was set up as follows: 10 ul Phusion HF Buffer 5x 1ul of dNTP 0.5 ul of 100 um primers 2 ul of AAV template 36 ul of water ................................................ 98 °C/45 s ................................................ (1x 98 °C/15 s 72 °C/30 s ................................................ (35x) 72 °C/10 min ................................................ 4 °C/ hold
File:Hot start 25102010 a.png fragments 1-8 of AAV1 and 1-4 of AAV2
File:Hot start 25102010 c.png fragments 1-8 of AAV8 and 1-4 of AAV5 26/10/2010
27/10/2010AAV 1 fragments 1-4; 5-8 and 1-8 (full capsid) assembly Touchdown Phusion HiFi PCR was performed according to the following protocol: ................................................ 98 °C/30s ................................................ (1x 98 °C/15 s 72 °C/15 s (- 1.0 °C/cycle) 72 °C/1 min ................................................ (17x) 98 °C/15 s 55 °C/30 s 72 °C/1 min ................................................ (20x) 72 °C/10 min ................................................ (1x) 4 °C/ hold ................................................
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||