Team:HokkaidoU Japan/Notebook/September3

From 2010.igem.org

(Difference between revisions)
(→前日のトラフォメの結果)
(→前日のトラフォメ菌のコロニーPCR)
Line 6: Line 6:
* colonies that should been red because of RFP insert wasn`t, so there is posibility that insert wasn`t there
* colonies that should been red because of RFP insert wasn`t, so there is posibility that insert wasn`t there
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=前日のトラフォメ菌の[[コロニーPCR]]=
+
=Colony PCR on yesterdays E.coli=
-
* プロトコル：コロニーPCRに従って，実験した
+
* Colony PCR was done acordig to protocol
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* 今回は20サンプルで行った
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* This day we did 20 samples
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===電気泳動===
+
 
 +
===Electrophoresis===
=前日に使用したパーツの濃度測定=
=前日に使用したパーツの濃度測定=

Revision as of 17:26, 27 September 2010

Notebook

Resultsof yesterdays trnsformation

  • pSB1C3 uterly failed to produce colonies
  • pUC119 produced 20 colonies
  • colonies that should been red because of RFP insert wasn`t, so there is posibility that insert wasn`t there

Colony PCR on yesterdays E.coli

  • Colony PCR was done acordig to protocol
  • This day we did 20 samples

Electrophoresis

前日に使用したパーツの濃度測定

HokkaidoU Japan 20100903a.jpg

Ligationに使用したDNA solutionが予想通りの濃度だったのか確認した

Lane DNA
1
2 λ/Hind III, EcoR I
3
4 RFP
5 pUC119
6 pSB1C3

1-3AのPCR

1-3AはpSB1C3に載ったRFP reporterで，トラフォメに成功している
ベクターとして配布されたpSB1C3が悪い可能性を見るため，このパーツのpSB1C3を増幅して使用する

Reagent Amount
1-3A 1
DW 33
10x Buffer 5
2 M 4dNTPs 5
25 mM MgSO4 3
Suffix-F 1
Prefix-R 1
KOD 1
Total 50 uL
  • extensionは120 sec
  • YM-10でClean Upしたら43 uLになった
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