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1 Notebook : August 9
- 1.1 Lab work
  - 1.1.1 A - Aerobic/Anaerobic regulation system
    - 1.1.1.1 Objective : obtaining Bba_K1155007
    - 1.1.1.2 1 - Extraction of Bba_K115007 from DH5α
  - 1.1.2 A - Aerobic/Anaerobic regulation system / B - PCB sensing system
    - 1.1.2.1 1 - Gibson assembly.

Notebook : August 9

Lab work

A - Aerobic/Anaerobic regulation system

((((((((((((((((((((====1 - Obtaining Δ fnr E. coli strain by transduction to test our biobricks====

Abdou, Anais, Damir, Nadia, XiaoJing

We do the exprience again because the lysis made on wensday didn't happen. It's probably because the phage strain was too old ( 2001)

Protocol : transduction

Picture: lysed cells comparison.

0µl phage(control): the petri dish is cloudy, bacteria are not lysed.
50µl phage: the petri dish is clear, bacteria are lysed by phages.
We used a wild type strain to keep a stock and a Δ fnr E. coli strain to obtain phages that might have encapsidated a Δfnr::Km fragment.

10µl,50µl and 100µl petri dishes are clear so phages are multiplied.

We let the antibiotic over night to select the right strain. ))))))))))))))))))))))

Objective : obtaining Bba_K1155007

1 - Extraction of Bba_K115007 from DH5α

Abdou

Protocol : Hight copy plamid extraction

We used colonies number 10, 14 and 15.

Nanodrop

Bba_K1155007 in clone 10 : 38ng/µl
Bba_K1155007 in clone 14: 48.5ng/µl
Bba_K1155007 in clone 15: 52 ng/µl

CONCLUSION !!!!!!!! We will sequence our plasmid.

A - Aerobic/Anaerobic regulation system / B - PCB sensing system

Objective : Obtaining FNR and BphR2 proteins (Gibson assembly)

1 - Gibson assembly.

August 1st PCR purification to be sure about the experiment. BphR2 part1, BphR2 part2 and RBS_BphR2_part1, FNR part1, FNR part2 and RBS_FNR_part1.

Protocol : PCR_clean_up

Results:

IMAGE

Well 1 : 6µL DNA Ladder
Well 2 : 5µL of BphR2 part1+1µl of 6X loading dy
Well 3 : 5µL of BphR2 part2+1µl of 6X loading dy
Well 4 : 5µL of RBS_BphR2 part1+1µl of 6X loading dy
Well 5 : 5µL of FNR part1+1µl of 6X loading dy
Well 6 : 5µL of FRN part2+1µl of 6X loading dy
Well 7: 5µL of RBS_FNR part1+1µl of 6X loading dy
Gel : 0.8%

we have no fragment so we must do again these PCRs

@@ Line 7: / Line 7: @@
 ==='''A - Aerobic/Anaerobic regulation system'''===
-===='''1 - Obtaining Δ ''fnr E. coli'' strain by transduction to test our biobricks'''====
+((((((((((((((((((((===='''1 - Obtaining Δ ''fnr E. coli'' strain by transduction to test our biobricks'''====
 Abdou, Anais, Damir, Nadia, XiaoJing
@@ Line 26: / Line 26: @@
 µl,50µl and 100µl petri dishes are clear so phages are multiplied.
-We let the antibiotic over night to select the right strain.
+We let the antibiotic over night to select the right strain. ))))))))))))))))))))))
+===='''Objective : obtaining Bba_K1155007'''====
+====1 - Extraction of Bba_K115007 from DH5α====
-===='''2 - Obtaining RBS_lacZ_ter. '''====
 Abdou
-Clone 10,14 and 15 plamid extraction using nucleospin kit.
+Protocol : [[Team:Paris_Saclay/Protocols/Hight copy plamid extraction|Hight copy plamid extraction]]
-Protocol : [[Team:Paris_Saclay/Protocols/hight copy plamid extraction|hight copy plamid extraction]]
+We used colonies number 10, 14 and 15.
+Nanodrop
+* Bba_K1155007 in clone 10 : 38ng/µl
+* Bba_K1155007 in clone 14: 48.5ng/µl
+* Bba_K1155007 in clone 15: 52 ng/µl
+{|
+| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" |
+CONCLUSION !!!!!!!! We will sequence our plasmid.
+|}
-Results: concentration measured by nanodrop
-*clone 10: C=38ng/µl   260/280= 1.78
-*clone 14: C=48.5ng/µl   260/280=1.90
-*clone 15: C=52 ng/µl     260/280=1.78
-We still have to sequence plasmids in order to verify our results.
 ==='''A - Aerobic/Anaerobic regulation system / B - PCB sensing system'''===

Team:Paris Saclay/Notebook/August/9

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