Team:Nanjing-China/Notebook

From 2013.igem.org

(Difference between revisions)

Revision as of 15:38, 16 September 2013

Home	Team	Official Team Profile	Project	Parts Submitted to the Registry	Modeling	Protocol	Safety	Attributions

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 {| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"
 !align="center"|[[Team:Nanjing-China|Home]]
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 !align="center"|[[Team:Nanjing-China/Parts|Parts Submitted to the Registry]]
 !align="center"|[[Team:Nanjing-China/Modeling|Modeling]]
-!align="center"|[[Team:Nanjing-China/Notebook|Notebook]]
+!align="center"|[[Team:Nanjing-China/Protocol|Protocol]]
 !align="center"|[[Team:Nanjing-China/Safety|Safety]]
 !align="center"|[[Team:Nanjing-China/Attributions|Attributions]]
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-You should make use of the calendar feature on the wiki and start a lab notebook.  This may be looked at by the judges to see how your work progressed throughout the summer.  It is a very useful organizational tool as well.
+===1. Extraction of mRNA===
+:Kit
+===2. RT-PCR===
+:Kit
+===3.PCR===
+====(1) Colony PCR====
+:{|border=0
+|width="125"|2×PrimeStar
+|width="125"|12.5μl
+|-
+|Forward primer||1~2.5μl
+|-
+|Reverse primer||1~2.5μl
+|-
+|Bacteria solution||1~2μl
+|-
+|dd water||up to 25μl
+|}
+:{|border=0
+|width="75"|Step 1
+|width="200"|94℃
+|-
+|Step 2||94℃
+|-
+|Step 3||Annealing temperature
+|-
+|Step 4||72℃
+|-
+|Step 5||goto step2, 25~35 cycles
+|-
+|Step 6||72℃
+|-
+|Pause at 4℃||
+|}
+====(2) PCR with DNA as template====
+:{|border=0
+|width="125"|2×PrimeStar
+|width="125"|12.5μl
+|-
+|Forward primer||1μl
+|-
+|Reverse primer||1μl
+|-
+|Template DNA||0.5μl
+|-
+|dd water||up to 25μl
+|}
+:{|border=0
+|width="75"|Step 1
+|width="200"|94℃
+|-
+|Step 2||94℃
+|-
+|Step 3||Annealing temperature
+|-
+|Step 4||72℃
+|-
+|Step 5||goto step2, 25~35 cycles
+|-
+|Step 6||72℃
+|-
+|Pause at 4℃||
+|}
+:Gel running and purify the product.

2×PrimeStar	12.5μl
Forward primer	1~2.5μl
Reverse primer	1~2.5μl
Bacteria solution	1~2μl
dd water	up to 25μl

Step 1	94℃
Step 2	94℃
Step 3	Annealing temperature
Step 4	72℃
Step 5	goto step2, 25~35 cycles
Step 6	72℃
Pause at 4℃

2×PrimeStar	12.5μl
Forward primer	1μl
Reverse primer	1μl
Template DNA	0.5μl
dd water	up to 25μl

Step 1	94℃
Step 2	94℃
Step 3	Annealing temperature
Step 4	72℃
Step 5	goto step2, 25~35 cycles
Step 6	72℃
Pause at 4℃

Team:Nanjing-China/Notebook

From 2013.igem.org

Revision as of 15:38, 16 September 2013

Contents

1. Extraction of mRNA

2. RT-PCR

3.PCR

(1) Colony PCR

(2) PCR with DNA as template