Team:HokkaidoU Japan/Notebook/August2

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==サンプルの調製==
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==Restriction enzyme digestion==
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# 乾燥DNAに30 uLのTEを加えた（DNA solution）
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Added 30 uL of TE into the DNA solution (purified via miniprep)
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# 次の組成で泳動サンプルを調製した
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===Digestion Menu===
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{| class="main" style="text-align:center"
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{| 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;"|
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|style="border-bottom:1px solid #000;"| サンプル１
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|style="border-bottom:1px solid #000;"| sample 1
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|style="border-bottom:1px solid #000;"| サンプル２
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|style="border-bottom:1px solid #000;"| sample 2
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|style="border-bottom:1px solid #000;"| サンプル３
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|style="border-bottom:1px solid #000;"| sample 3
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|style="border-bottom:1px solid #000;"| サンプル４
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|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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# これらのサンプルを調製後，37℃で1時間おいた
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# incubated at 37C for 1 hrs
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# 各サンプルに2 uLの6x sample bufferを加えて混ぜた
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# add 2 uL of 6x sample buffer
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# 電気泳動
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# Electrophoresed
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==電気泳動==
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==Result of Electrophoresis==
[[Image:HokkaidoU Japan 20100802b.jpg‎|200px|right|thumb|]]
[[Image:HokkaidoU Japan 20100802b.jpg‎|200px|right|thumb|]]
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* 制限酵素で切り出したパーツ部分は1069 bp（＋制限サイト
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* The insert lenght wants 1069 bp (plus RE recognition site, 49 bp)
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* Plasmid backboneは5263 bp
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* Plasmid backbone was 5263 bp
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* マーカーはλ/''Hin''d IIIとpUC119/''Hin''f I
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* Marker was λ/''Hin''d III and pUC119/''Hin''f I
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Good

Latest revision as of 06:57, 27 October 2010

Notebook

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


  1. incubated at 37C for 1 hrs
  2. add 2 uL of 6x sample buffer
  3. Electrophoresed

Result of Electrophoresis

HokkaidoU Japan 20100802b.jpg
  • 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
Good
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