Team:UNIPV-Pavia/Calendar/August/settimana2
From 2010.igem.org
m (→August, 9th) |
(→August, 9th) |
||
Line 38: | Line 38: | ||
|} | |} | ||
In order not to loose our time we decided to perform again the PCR-amplification/modification of <partinfo>BBa_K208001</partinfo> in case of a negative screening. Than we gel-ran and gel-extracted right amplicons. | In order not to loose our time we decided to perform again the PCR-amplification/modification of <partinfo>BBa_K208001</partinfo> in case of a negative screening. Than we gel-ran and gel-extracted right amplicons. | ||
+ | PCR was performed with: | ||
+ | *10_F and S_R primers | ||
+ | *S_F and S_R primers | ||
+ | [[Image:UNIPV10_BW23474_J72007.jpg|thumb|200px|center|BW23474 transformed with <partinfo>BBa_J72007</partinfo>]] | ||
+ | |||
+ | The image shows results of our PCR: all 2 parts are positive, so we purified cut DNA, obtaining the following quantifications: | ||
+ | *PhaP 10-S: 16,8 ng/ul | ||
+ | *PhaP S-S: 17,1 ng/ul | ||
+ | |||
+ | Tomorrow we will proceed to digest this part, in order to have an alternative if all clones of I20 and I21 are negative. | ||
+ | |||
+ | In the afternoon, we inoculated all parts we will need tomorrow. | ||
+ | |||
+ | {| border='1' align='center | ||
+ | |''Part'' || ''Medium'' || ''Task'' | ||
+ | |- | ||
+ | |<partinfo>BBa_B0034</partinfo> || 5ml LB+Amp || Ligation | ||
+ | |- | ||
+ | |I15-1 || 5ml LB+Amp || Ligation | ||
+ | |- | ||
+ | | Linker || 5ml LB+Amp || Ligation | ||
+ | |- | ||
+ | | pSB4C5 || 5ml LB+Cm 12,5 || Ligation | ||
+ | |- | ||
+ | | I22-1 || 5ml LB+ Amp (picked from colony)|| Ligation/Screening/Glycerol stock | ||
+ | |- | ||
+ | | I22-2 || 5ml LB+ Amp (picked from colony)|| Ligation/Screening/Glycerol stock | ||
+ | |- | ||
+ | | I22-3 || 5ml LB+ Amp (picked from colony)|| Ligation/Screening/Glycerol stock | ||
+ | |- | ||
+ | | I23-1 || 5ml LB+ Amp (picked from colony)|| Screening/Glycerol stock | ||
+ | |- | ||
+ | | I23-2 || 5ml LB+ Amp (picked from colony)|| Screening/Glycerol stock | ||
+ | |- | ||
+ | | I23-3 || 5ml LB+ Amp (picked from colony)|| Screening/Glycerol stock | ||
+ | |- | ||
+ | | <partinfo>BBa_r0062</partinfo> || 5ml LB+ Amp|| Ligation | ||
+ | |- | ||
+ | | I7_4C5-2 || 1ml LB+ Cm12,5 || Tecan Test | ||
+ | |- | ||
+ | | I8_4C5-2 || 1ml LB+ Cm12,5 || Tecan Test | ||
+ | |- | ||
+ | | I10_4C5-1 || 1ml LB+ Cm12,5 || Tecan Test | ||
+ | |- | ||
+ | | I12_4C5-1 || 1ml LB+ Cm12,5 || Tecan Test | ||
+ | |- | ||
+ | | A2(=<partinfo>BBa_J23100</partinfo>+GFP) || 1ml LB+ Amp || Tecan Test | ||
+ | |- | ||
+ | | <partinfo>BBa_B0032</partinfo>|| 1ml LB+ Amp || Tecan Test | ||
+ | |- | ||
+ | | I8-5 D || 1ml LB+ Amp (picked from colony)|| Tecan Test/Glycerol stock | ||
+ | |- | ||
+ | | I8-5 E || 1ml LB+ Amp (picked from colony)|| Tecan Test/Glycerol stock | ||
+ | |- | ||
+ | | I8-5 F || 1ml LB+ Amp (picked from colony)|| Tecan Test/Glycerol stock | ||
+ | |} | ||
---- | ---- | ||
Check of LB+Cm 6 ug/ml agar plates. We let grow at 30°C: | Check of LB+Cm 6 ug/ml agar plates. We let grow at 30°C: |
Revision as of 13:32, 10 August 2010
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|