Team:HokkaidoU Japan/Notebook/August24

From 2010.igem.org

(Difference between revisions)
(→Ligation Mixtureのチェック)
(→いろいろ確認)
Line 36: Line 36:
* 100 uL of each solution was plated on mediums containing 34 ug/uL and 12 ug/uL of chloramphenicol
* 100 uL of each solution was plated on mediums containing 34 ug/uL and 12 ug/uL of chloramphenicol
-
=いろいろ確認=
+
=Various Checks=
[[Image:HokkaidoU Japan 20100824b.jpg‎|200px|right|thumb|]]
[[Image:HokkaidoU Japan 20100824b.jpg‎|200px|right|thumb|]]
{| class="protocol"
{| class="protocol"
Line 43: Line 43:
|-
|-
|2
|2
-
|λ/''Hin''d III
+
|[https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png Lambda/''Hin''d III]
|-
|-
|3
|3
-
|pSB1C3どうしのLigation
+
|Ligation between pSB1C3 vectors
|-
|-
|4
|4
-
|ラオリプライマーによる1-2NのPCRのろ過産物（上）
+
|Product obtained by using digestion visualization primer for 1-2N
|-
|-
|5
|5
-
|ラオリプライマーによる1-2NのPCRのろ過（下）
+
|Flow through of digestion visualization primer for 1-2N
|-
|-
|6
|6
-
|λ/''Hin''d III
+
|[https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png Lambda/''Hin''d III]
|-
|-
|7
|7
-
|ゲル抽したpSB1C3
+
|Gel extracted pSB1C3
|}
|}
-
レーン３でバンドが見られなかった．
+
Band weren't visible in lane 3
-
* ダイマー，テトラマーなど，さまざまな断片ができ，薄くなってしまった？
+
* Dimers, trimers and etc were hardly visible maybe severely diluted
-
レーン７は薄いがバンドが確認できた
+
 
-
* ゲル抽に問題はない
+
In lane 7 had detectable by very weak band
 +
* There weren't any problems with gel extraction

Revision as of 09:01, 22 September 2010

Notebook

Check to see if digestion visualization Primers Work

Electrophoresis after restriction enzyme digestion‎
  • All 4 of PCR products were purified via Microcon YM-10
Lane DNA
1 TSUDA Marker I
3 RBS
4 GFP
5 double terminator
6 Promoter

Promoter band in lane 6 wasn't visible

  • This vector didn't have a primer annealing site for our new primers

Check of Ligation Mixtures

TAKARABIO's Ligation Mixture [I] and Ligation Mixture Mighty Mix [M]

  • After PCR and gel extracted pSB1C3 volume of either Ligation Mixture equal to pSB1C3 solution volume was added
  • Transformed
  • incubated in 200 uL of LB medium
  • 100 uL of each solution was plated on mediums containing 34 ug/uL and 12 ug/uL of chloramphenicol

Various Checks

HokkaidoU Japan 20100824b.jpg
Lane DNA
2 Lambda/Hind III
3 Ligation between pSB1C3 vectors
4 Product obtained by using digestion visualization primer for 1-2N
5 Flow through of digestion visualization primer for 1-2N
6 Lambda/Hind III
7 Gel extracted pSB1C3

Band weren't visible in lane 3

  • Dimers, trimers and etc were hardly visible maybe severely diluted

In lane 7 had detectable by very weak band

  • There weren't any problems with gel extraction
Retrieved from "http://2010.igem.org/Team:HokkaidoU_Japan/Notebook/August24"