IGEM:Peking University/2008/Notebook/Group 1/2008/07/18: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
(Autocreate 2008/07/18 Entry for IGEM:Peking_University/2008/Notebook/Group_1) |
|||
Line 5: | Line 5: | ||
|- | |- | ||
| colspan="2"| | | colspan="2"| | ||
== | ==PKU_080718_G1_1== | ||
<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
Group 1 | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
PKU_080718_G1_1
|