IGEM:Peking University/2008/Notebook/Group 3/2008/07/29: Difference between revisions

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(Autocreate 2008/07/29 Entry for IGEM:Peking_University/2008/Notebook/Group_3)
 
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| colspan="2"|
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==Entry title==
==PKU_080729_G3_1==
* Insert your content here.
 
 
::'''Enzyme cutting-testing-pGREG503-lacI'''
 
System1:''Giving up cause no EcoRI''
 
 
System2:
 
{|border="1" cellspacing="0" align="center"
|+
|-
| ||Volume (μL)
|-
| pGREG503-lacI||5
|-
|SacI||0.2×22
|-
|XbaI||0.2
|-
|10×M Buffer||2
|-
|ddH<sub>2</sub>O||12.6
|}
 
Result: false positive transformation
 
 
<hr class = divider>
 
 
::'''PCR system'''
{|border="1" cellspacing="0" align="center"
|+
|-
| ||Volume (μL)
|-
|10×PCR buffer||2||×22
|-
| transtart||0.1
|-
|dNTP||2
|-
|template||2
|-
|F-Primer||0.4
|-
|R-Primer||0.4
|-
|ddH<sub>2</sub>O||13.1
|}
 
 
::'''conditions'''
{|border="1" cellspacing="0" align="center"
|+
|-
|Temperature(°C)||Time Length(0:00:00)
|-
| 94||02:00
|-
| 94||00:20||30 cycles
|-
| 60||00:30||30 cycles
|-
| 72||07:00
|}
 
 
Reinforce the result of false positive transformation
 
 
<hr class = divider>
 
 
::'''Colony PCR'''
 
 
{|border="1" cellspacing="0" align="center"
|+
|-
| ||Volume (μL)
|-
|Buffer ||1.5||×20
|-
|Easy tag||0.1
|-
|dNTP||1.5
|-
|primer-R||0.5
|-
|primer-F||0.5
|-
|ddH<sub>2</sub>O||11
|}
 
 
 




Revision as of 05:54, 29 July 2008

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PKU_080729_G3_1

Enzyme cutting-testing-pGREG503-lacI

System1:Giving up cause no EcoRI


System2:

Volume (μL)
pGREG503-lacI 5
SacI 0.2×22
XbaI 0.2
10×M Buffer 2
ddH2O 12.6

Result: false positive transformation



PCR system
Volume (μL)
10×PCR buffer 2 ×22
transtart 0.1
dNTP 2
template 2
F-Primer 0.4
R-Primer 0.4
ddH2O 13.1


conditions
Temperature(°C) Time Length(0:00:00)
94 02:00
94 00:20 30 cycles
60 00:30 30 cycles
72 07:00


Reinforce the result of false positive transformation



Colony PCR


Volume (μL)
Buffer 1.5 ×20
Easy tag 0.1
dNTP 1.5
primer-R 0.5
primer-F 0.5
ddH2O 11





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