"
Team:Heidelberg/Templates/DelH week25
From 2013.igem.org
(Difference between revisions)
| Line 1: | Line 1: | ||
| - | + | =18.10= | |
==HM17-FS_78== | ==HM17-FS_78== | ||
Annealing temps: 62°C, 60°C, 57°C, 55°C <br/> | Annealing temps: 62°C, 60°C, 57°C, 55°C <br/> | ||
| Line 28: | Line 28: | ||
12 cycles 74, 68, 64 and 58°C <br/> | 12 cycles 74, 68, 64 and 58°C <br/> | ||
18 cycles 72, 66, 62 and 56°C | 18 cycles 72, 66, 62 and 56°C | ||
| + | |||
| + | =20-10-2013= | ||
| + | ==Inocultation triple dummy clone== | ||
| + | ==Screening triple clone== | ||
| + | each with positive control | ||
| + | IK25+VF2 -->54°C, 90s elongation <br/> | ||
| + | FS47+FS48 --> 66t,62 40s elongation <br/> | ||
| + | FS95+FS73 --> 66t,62 40s elongation <br/> | ||
Revision as of 15:49, 20 October 2013
Contents |
18.10
HM17-FS_78
Annealing temps: 62°C, 60°C, 57°C, 55°C
Elongation time: 2:10
--> Did not work
Screening remaining Gibson electroporated
60 colonies were picked and screened with primers FS_73 and FS_95 (annealing 66 touchdown, 62)
Gibson assembly
| Reagent | Volume added (µl) |
|---|---|
| BB (FS..) | 0.61 |
| DN11-FS66 | 1.00 |
| FS_67-FS_68 | 1.35 |
| FS_69-FS_70 | 6.34 |
| FS_71-HM08 | 0.70 |
| Gibson Master Mix 2x | 10 |
ccdB
Annealing temps: </br>
12 cycles 74, 68, 64 and 58°C
18 cycles 72, 66, 62 and 56°C
20-10-2013
Inocultation triple dummy clone
Screening triple clone
each with positive control
IK25+VF2 -->54°C, 90s elongation
FS47+FS48 --> 66t,62 40s elongation
FS95+FS73 --> 66t,62 40s elongation
