Team:Heidelberg/Notebook/miMeasure/July
From 2010.igem.org
(→05/07/2010 - 11/07/2010) |
(→12/07/2010 - 19/07/2010) |
||
Line 8: | Line 8: | ||
Cell Culture Starting | Cell Culture Starting | ||
- | = 12/07/2010 - 19/07/2010 = | + | ===12/07/2010 - 19/07/2010=== |
<br /> | <br /> | ||
- | + | Preperation of RNA extracts for miRNA profiling | |
<br /> | <br /> | ||
*Hela, Hek and HUH were plated on p100 dishes in their according media (10E6 cells/dish) and grown to 60-70 % confluence | *Hela, Hek and HUH were plated on p100 dishes in their according media (10E6 cells/dish) and grown to 60-70 % confluence | ||
Line 20: | Line 20: | ||
*pSB1AC3 from the registry was transformed into Top10 cells according to the standard transformation protocol; a day later, a 5 ml LB culture was inocculated and incubated for 8 hours; plasmid was extracted by applying the Qiagen MiniPrep Protocol | *pSB1AC3 from the registry was transformed into Top10 cells according to the standard transformation protocol; a day later, a 5 ml LB culture was inocculated and incubated for 8 hours; plasmid was extracted by applying the Qiagen MiniPrep Protocol | ||
<br /> | <br /> | ||
- | + | design of diraPCR oligos | |
<br /> | <br /> | ||
* a diraPCR Designer tool (called miRACLE Designer) was programmed for enabling quick&easy design of diraPCR oligos. The program takes any microRNA guiding strand sequence of choice and constructs synthetic microRNA binding site oligos. Those oligos have the following properties: | * a diraPCR Designer tool (called miRACLE Designer) was programmed for enabling quick&easy design of diraPCR oligos. The program takes any microRNA guiding strand sequence of choice and constructs synthetic microRNA binding site oligos. Those oligos have the following properties: | ||
Line 30: | Line 30: | ||
<br /> | <br /> | ||
- | + | design of measurement standard | |
<br /> | <br /> | ||
* in order to design an appropriety standard respecting all the rules according to the standard of mammalian synthetic biology (RFC12), we developed a dual reporter measurment construct with the following properties: | * in order to design an appropriety standard respecting all the rules according to the standard of mammalian synthetic biology (RFC12), we developed a dual reporter measurment construct with the following properties: |
Revision as of 10:43, 26 October 2010
|
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||