Team:Tokyo Metropolitan/Notebook/Fiber/2010/08/17

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Latest revision as of 21:38, 26 October 2010

E.coli Fiber Project Notebook

2010/8/17 Tuesday (watachin)

Experiment:Electrophoresis

Member

NEX and watachin

Material

pSB1A3(25ng/μl) 22μl
10×Loading buffer 2.2μl
DNA Marker 5μl
1×TAE buffer
1% agarose gel

Procedure

Set agarose gel and add TAE buffer in electrophoresis tank.
Mix Loading Buffer and pSB1C3 ,then put them in well (marker sets another well).
Load DNA at 100V for two thirds of total volume (about 15 minutes).
Image the consequence of electrophoresis.

Result

failure

Plasmid concentration was too low.

Experiment:Transformation of pSB1A3

Member

NEX and watachin

Material

pSB1A3 1μl
competent cell (DH5α) 50μl
LB + amp plate

Procedure

mix pSB1A3 and DH5α
on ice (30min)
heat shock 42℃ (45sec)
on ice (2min)
inoculate onto plate
incubate cells at 37℃

@@ Line 2: / Line 2: @@
 ==2010/8/17 Tuesday (watachin)==
-===Experiment:Electrophoreses of PCR productions===
+===Experiment:Electrophoresis===
-'''Member'''<br />
+:'''Member'''
-NEX and watachin
+:NEX and watachin
-'''Materials'''
+:'''Material'''
-*pSB1A3(25ng/μl) 22μl
+:*pSB1A3(25ng/μl) 22μl
-*10*Loading buffer 2.2μl
+:*10×Loading buffer 2.2μl
-*DNA Marker 5μl
+:*DNA Marker 5μl
-*1*TAE buffer
+:*1×TAE buffer
-*1% agarose gel
+:*1% agarose gel
-'''Procedure'''
+:'''Procedure'''
-#Set agarose gel and add TAE buffer in electrophoresis tank.
+:#Set agarose gel and add TAE buffer in electrophoresis tank.
-#Mix Loading Buffer and pSB1C3 ,then put them in well (marker sets another well).
+:#Mix Loading Buffer and pSB1C3 ,then put them in well (marker sets another well).
-#Load DNA at 100V for two thirds of total volume (about 15 minutes).
+:#Load DNA at 100V for two thirds of total volume (about 15 minutes).
-#Image the consequence of electrophoresis.
+:#Image the consequence of electrophoresis.
-'''Result'''
+:'''Result'''
 [[Image:2010-08-17-ef.jpg]]
-failure
+:failure
-Plasmid concentration was too low.
+:Plasmid concentration was too low.
 ===Experiment:Transformation of pSB1A3===
-'''Member'''<br />
+:'''Member'''
-NEX and watachin
+:NEX and watachin
-'''Material'''
+:'''Material'''
-*pSB1A3 1μl
+:*pSB1A3 1μl
-*competent cell DH5α 50μl
+:*competent cell (DH5α) 50μl
-*LB + amp plate
+:*LB + amp plate
-'''Procedure'''
+:'''Procedure'''
-#mix pSB1A3 and DH5α
+:#mix pSB1A3 and DH5α
-#on ice (30min)
+:#on ice (30min)
-#heat shock 42℃ 45sec
+:#heat shock 42℃ (45sec)
-#on ice (2min)
+:#on ice (2min)
-#inoculate this onto plate
+:#inoculate onto plate
-#incubate cells at 37℃
+:#incubate cells at 37℃
+<br/>