Team:HokkaidoU Japan/Notebook/September3

From 2010.igem.org

(Difference between revisions)
(→前日のトラフォメ菌のコロニーPCR)
(→前日に使用したパーツの濃度測定)
Line 12: Line 12:
===Electrophoresis===
===Electrophoresis===
-
=前日に使用したパーツの濃度測定=
+
=Concentration check of parts used for transformation yesterday=
-
[[Image:HokkaidoU_Japan_20100903a.jpg|200px|right|thumb|]]
+
 
-
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
+
[[Image:HokkaidoU_Japan_20100903a.jpg|200px|right|thumb|Electrophoresis of parts before ligation]]
 +
 
 +
Confirmed that DNA solution concentration of parts for Ligation was as anticipated (good?)
 +
 
{|class="protocol"
{|class="protocol"
|-
|-
Line 24: Line 27:
|-
|-
|2
|2
-
|λ/Hind III, EcoR I
+
|[https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png Lambda/Hind III, EcoR I]
|-
|-
|3
|3

Revision as of 17:29, 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

Concentration check of parts used for transformation yesterday

Electrophoresis of parts before ligation

Confirmed that DNA solution concentration of parts for Ligation was as anticipated (good?)

Lane DNA
1
2 Lambda/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
Retrieved from "http://2010.igem.org/Team:HokkaidoU_Japan/Notebook/September3"