Revision as of 10:01, 22 September 2010

Transformation Results

1-3A had colonies in lated on mediums containing 34 ug/uL of chloramphenicol
- So the medium and antibiotic consentration seems ok,
Medium on which M 12 was plated was inoculated with chloramphenicol after solidification so there were some irregularities and colonies on the fringe
Colonies of 1-3A and 12 were obviously different

→ So we suspected issues in Ligation and DNA concentration bused for transformation

Electrophoresis of 1uL of 1-3A DNA

→Estimated concentration 2 ng/uL

Tomorrow will do aditional Electrophoresis gel extraction and transformation

前日のPCRから，増やすことのできたパーツのDNA量を求めた

→37℃で60分

→ラオリプライマーの真価を電気泳動で確認

@@ Line 43: / Line 43: @@
 <div style="clear:both"></div>
-=pSB1C3をエタ沈で濃縮，Ligation=
+=Ethanol precipitation of pSB1C3 and Ligation=
-===エタ沈===
-# 3 M 酢酸ナトリウム 8.2 uL加える
+===Ethanol precipitation===
-# 100%エタノールを205 uL加え，混ぜる
-# 15,00rpm@4℃で10分間遠心する
+# Added 8.2 uL of sodium acetate [3 M]
-# 上清を捨て，500 uL 70%エタノールを加え，混ぜる
+# Added 205 uL of Ethanol [100%] and mixed
-# 15,00rpm@4℃で5分間遠心する
+# Centrifuge at 15,00rpm, 4C for 10 min
-# 上清を捨て，真空デシケータで乾燥させる
+# Discarded supernatant and added 500 uL of Ethanol [70%], mixed
-# DW 8.2 uLを加え，元の10倍に濃縮する
+# Centrifuge at 15,00rpm, 4C for 5 min
+# Discarded supernatant
+# Dried via vacuum desiccator
+# Added 8.2 uL of ADW を加え，元の10倍に濃縮する
+* Concentration increased 10 fold
 ===Ligation===
-# 8.2 uL DNA solutionに同量のLigation Solutionを加える
+# Mixed 8.2 uL of DNA solution with the same volume of Ligation Solution
-# T4 ligase 1 uLを加え，16℃で30分間インキュベート
+# Added extra 1 uL of T4 ligase
-# 明日，電気泳動・ゲル抽・Transformation
+# Incubated at 16C for 30 min
+Tomorrow will do aditional Electrophoresis gel extraction and transformation
 =らおりプライマーで作ったPCR productsのDigestion=