"
30/07/13
From 2013.igem.org
(Difference between revisions)
TanviSinha (Talk | contribs) |
(→Transformation of the limonene Biobrick (BBa_K118025)) |
||
| (One intermediate revision not shown) | |||
| Line 1: | Line 1: | ||
| - | {| style="color:# | + | <div style="font-family:arial;padding:5px;border-radius:5px;border:5px solid #FF2800;"> |
| + | {| style="color:#87EA00;background-color:#FFFFFF;" cellpadding="2" cellspacing="2" border="0" bordercolor="#000000" width="100%" align="center" | ||
!align="center"|[[Team:Leicester|Home]] | !align="center"|[[Team:Leicester|Home]] | ||
!align="center"|[[Team:Leicester/Team|Team]] | !align="center"|[[Team:Leicester/Team|Team]] | ||
| Line 10: | Line 11: | ||
!align="center"|[[Team:Leicester/Attributions|Attributions]] | !align="center"|[[Team:Leicester/Attributions|Attributions]] | ||
|} | |} | ||
| + | </div> | ||
<p> | <p> | ||
| Line 16: | Line 18: | ||
==Transformation of the limonene Biobrick (BBa_K118025)== | ==Transformation of the limonene Biobrick (BBa_K118025)== | ||
Ligated with the pSB1C3 vector (restriction sites EcoRI, PstI) | Ligated with the pSB1C3 vector (restriction sites EcoRI, PstI) | ||
| - | |||
<b>Protocol:</b> | <b>Protocol:</b> | ||
Latest revision as of 14:37, 2 August 2013
| Home | Team | Official Team Profile | Project | Parts Submitted to the Registry | Modeling | Notebook | Safety | Attributions |
|---|
Transformation of the limonene Biobrick (BBa_K118025)
Ligated with the pSB1C3 vector (restriction sites EcoRI, PstI)
Protocol:
- Start thawing the competent cells on ice
- Seperate 4 pre-chilled eppendorf tubes;
- Resistance/Viability test
- Ligation experiment with 10ul DNA
- Ligation experiment with 5ul DNA
- Positive control using 1ul RFP
- Add 50ul of thawed competent cells and then the aforementioned volume of DNA to each eppendorf tube.
- For each eppendorf tube, pipette the solution gently to mix. Ensure the cells are kept on ice.
- Close the tubes and incubate the cells on ice for 30 minutes
- Heat shock the cells by immersion in a pre-heated water bath at 42'C for 60 seconds
- Incubate the cells on ice for 5 minutes
- Add the 200ul of SOC (SOB + 0.4g glucose) media to each tube
- Incubate the cells at 37'C for 2 hours
- Plate onto petri dishes (ensure your petri dishes are labelled correctly):
- For eppendorf 1, plate 100ul onto a chloroamphenicol petri dish and another 100ul of the solution onto a LB (luria broth) petri dish
- For eppendorf 2, plate 200ul and 20ul onto two separate chloroamphenicol petri dishes
- For eppendorf 3, plate 200ul and 20ul onto two separate chloroamphenicol petri dishes
- For eppendorf 4, plate 200ul and 20ul onto two separate chloroamphenicol petri dishes
- Incubate the plates at 37'C overnight
