Revision as of 11:56, 11 August 2013

Notebook : August 9

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)

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.

Now we can do a gel purification of the highest band which contains the PSB3K3 plasmid.

2.2 - Electroelution of the highest band to extract the PSB3K3 plasmid from the gel

Nadia

We let the plasmid precipitate during the night.

Anaïs

Colony counting :
- Low concentration petri dish : 47 colonies
- High concentration petri dish : 145 colonies

Protocol : Colony PCR

PCR Program :
 [IMAGE]

Anaïs, Damir

Expected size : 3583bp

Colonies 10, 14, 15 exhibit plasmids with the right length.

@@ Line 18: / Line 18: @@
 | style="width:350px;border:1px solid black;" | [[File:Ps908transduction.jpg|350px]]
 | style="width:350px;border:1px solid black;vertical-align:top;" |
-*Well 1 : 6µL DNA Ladder
+'''picture: lysed cells comparison.'''
-*Well 2 : 45µL of BBa_J004450 digested by EcoRI/Pst1
+* 0µl phage(control): the petri dish is cloudy, bacteria are not lysed
-*Gel : 0.8%
+* 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.
 |}