IGEM:Peking University/2008/Notebook/Group 1/2008/07/18: Difference between revisions

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(Autocreate 2008/07/18 Entry for IGEM:Peking_University/2008/Notebook/Group_1)
 
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==Entry title==
==PKU_080718_G1_1==
* Insert your content here.
 
<hr class=divider>
::'''Enzyme cutting-pADH-lac2(PCR product)'''
 
::::::::::'''System for double ligation'''
 
{|border="1" cellspacing="0" align="center"
|+
|-
| ||Volume (μL)
|-
| Promoter||20
|-
|BamHI||1
|-
|SacI||1
|-
|0.5×K Buffer||2.5
|-
|ddH<sub>2</sub>O||25.5
|}
 
::::::::::'''System for single ligation'''
 
{|border="1" cellspacing="0" align="center"
|+
|-
| ||Volume (μL)
|-
| Promoter||20
|-
|XhoI||2
|-
|SacI||2
|-
|1×M Buffer||5
|-
|ddH<sub>2</sub>O||21
|}
 
 
<hr class=divider>
::'''PCR- pGREG504-pADH-(lac1/lacS/tet)-50ul''
::''Using program lac12MQT''
 
{|
[[Image:PKU-08717-PCR-pADH-promoters.tif|center|400px]]
|}
 
<hr class=divider>
::'''Gel running and extraction-promoter(PCR product)'''
 
<hr class=divider>
::'''Enzyme cutting-AID'''
 
{|
[[Image:PKU-080718 lac2(double single) AID digestion.tif|center|400px]]
|}
 
:::::::'''lanes: intact AID(control)|λmarker|Promoter(single)|promoter(double)|AID|AID'''
 
::::::::::'''System for double ligation'''
 
{|border="1" cellspacing="0" align="center"
|+
|-
| ||Volume (μL)
|-
| Promoter||20
|-
|BamHI||1||×3
|-
|SacI||1
|-
|0.5×K Buffer||2.5
|-
|ddH<sub>2</sub>O||25.5
|}
 
::::::::::'''System for single ligation'''
 
{|border="1" cellspacing="0" align="center"
|+
|-
| ||Volume (μL)
|-
| Promoter||20
|-
|XhoI||2||×3
|-
|SacI||2
|-
|1×M Buffer||5
|-
|ddH<sub>2</sub>O||21
|}
 
 
<hr class=divider>
::'''Single Ligation-pGREG504-pAID-lac2'''
{|border="1" cellspacing="0" align="center"
|+
|-
| ||Volume (μL)
|-
| lac2 ||4
|-
| pGREG504 ||1
|-
| ligation buffer||5
|}
 
 
::'''Double Ligation-pGREG504-pAID-lac2-AID'''
 
{|border="1" cellspacing="0" align="center"
|+
|-
| ||Volume (μL)
|-
| lac2 ||3
|-
| AID ||3
|-
| pGREG504 ||1
|-
| ligation buffer||7
|}
 
 
<hr class=divider>
::'''Gel running-testing-pADH-lac1/S/tet'''
::'''Gel running-testing-digestion-lac2/AID'''




Revision as of 08:40, 18 July 2008

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PKU_080718_G1_1

Enzyme cutting-pADH-lac2(PCR product)
System for double ligation
Volume (μL)
Promoter 20
BamHI 1
SacI 1
0.5×K Buffer 2.5
ddH2O 25.5
System for single ligation
Volume (μL)
Promoter 20
XhoI 2
SacI 2
1×M Buffer 5
ddH2O 21


'PCR- pGREG504-pADH-(lac1/lacS/tet)-50ul
Using program lac12MQT
Gel running and extraction-promoter(PCR product)
Enzyme cutting-AID
lanes: intact AID(control)|λmarker|Promoter(single)|promoter(double)|AID|AID
System for double ligation
Volume (μL)
Promoter 20
BamHI 1 ×3
SacI 1
0.5×K Buffer 2.5
ddH2O 25.5
System for single ligation
Volume (μL)
Promoter 20
XhoI 2 ×3
SacI 2
1×M Buffer 5
ddH2O 21


Single Ligation-pGREG504-pAID-lac2
Volume (μL)
lac2 4
pGREG504 1
ligation buffer 5


Double Ligation-pGREG504-pAID-lac2-AID
Volume (μL)
lac2 3
AID 3
pGREG504 1
ligation buffer 7


Gel running-testing-pADH-lac1/S/tet
Gel running-testing-digestion-lac2/AID



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