Team:HokkaidoU Japan/Notebook/August2
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{{Template:HokkaidoU_Japan}} | {{Template:HokkaidoU_Japan}} | ||
- | + | <div class="linkbar"><div class="prev">[[Team:HokkaidoU_Japan/Notebook/July27|July 27]]</div>[[Team:HokkaidoU_Japan/Notebook|Notebook]]<div class="next">[[Team:HokkaidoU_Japan/Notebook/August10|August 10]]</div></div> | |
- | + | ==Restriction enzyme digestion== | |
- | + | Added 30 uL of TE into the DNA solution (purified via miniprep) | |
- | {|style="text-align:center" | + | ===Digestion Menu=== |
+ | {| class="protocol" style="text-align:center" | ||
|- | |- | ||
|style="border-bottom:1px solid #000; border-right:1px solid #000;"| | |style="border-bottom:1px solid #000; border-right:1px solid #000;"| | ||
- | |style="border-bottom:1px solid #000;"| | + | |style="border-bottom:1px solid #000;"| sample 1 |
- | |style="border-bottom:1px solid #000;"| | + | |style="border-bottom:1px solid #000;"| sample 2 |
- | |style="border-bottom:1px solid #000;"| | + | |style="border-bottom:1px solid #000;"| sample 3 |
- | |style="border-bottom:1px solid #000;"| | + | |style="border-bottom:1px solid #000;"| sample 4 |
|- | |- | ||
|style="border-right:1px solid #000;"| DNA solution | |style="border-right:1px solid #000;"| DNA solution | ||
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| 0.5 uL | | 0.5 uL | ||
|} | |} | ||
+ | |||
+ | |||
+ | # incubated at 37C for 1 hrs | ||
+ | # add 2 uL of 6x sample buffer | ||
+ | # Electrophoresed | ||
+ | |||
+ | ==Result of Electrophoresis== | ||
+ | [[Image:HokkaidoU Japan 20100802b.jpg|200px|right|thumb|]] | ||
+ | * The insert lenght wants 1069 bp (plus RE recognition site, 49 bp) | ||
+ | * Plasmid backbone was 5263 bp | ||
+ | * Marker was λ/''Hin''d III and pUC119/''Hin''f I | ||
+ | Good |
Latest revision as of 06:57, 27 October 2010
Restriction enzyme digestion
Added 30 uL of TE into the DNA solution (purified via miniprep)
Digestion Menu
sample 1 | sample 2 | sample 3 | sample 4 | |
DNA solution | 1 uL | 1 uL | 1 uL | 1 uL |
10x H Buffer | - | 1 uL | 1 uL | 1 uL |
DW | 9 uL | 7.5 uL | 7.0 uL | 7.5 uL |
Xba1 | - | 0.5 uL | 0.5 uL | - |
Pst1 | - | - | 0.5 uL | 0.5 uL |
- incubated at 37C for 1 hrs
- add 2 uL of 6x sample buffer
- Electrophoresed
Result of Electrophoresis
- The insert lenght wants 1069 bp (plus RE recognition site, 49 bp)
- Plasmid backbone was 5263 bp
- Marker was λ/Hind III and pUC119/Hinf I