Revision as of 15:52, 12 August 2013

Notebook : August 12

Obtaining Pfnr_RBS-LacZ-Term in PSB1C3

Anaïs, Nadia, XiaoJing

Protocol : digestion

Used quantities :

IMAGE

Abdou

Clone 10,14 and 15 plamid extraction using nucleospin kit.

Results: concentration measured by nanodrop

We still have to sequence plasmids in order to verify our results.

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.

Results:

IMAGE

we have no fragment so we must do again these PCRs

@@ Line 37: / Line 37: @@
 ** PstI : 1µL
-| style="width:350px;border:1px solid black;" | [[File:Ps908transduction.jpg|350px]]
+{|
+| style="width:350px;border:1px solid black;" | IMAGE
 | style="width:350px;border:1px solid black;vertical-align:top;" |
-'''Picture: lysed cells comparison.'''
+*Well 1 : 6µL DNA Ladder
-* 0µl phage(control): the petri dish is cloudy, bacteria are not lysed.
+*Well 2 : 5µL of digested Bba_K1155000+1µl of 6X loading dy
-* 50µl phage: the petri dish is clear, bacteria are lysed by phages.
+*Well 3 : 5µL of digested Bba_K1155007+1µl of 6X loading dy
-*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.
+*Well 4 : 5µL of digested Bba_K1155003+1µl of 6X loading dy
+*Well 5 : 5µL of FNR part1+1µl of 6X loading dy
+*Gel : 0.8%
 |}
-µl,50µl and 100µl petri dishes are clear so phages are multiplied.
-We let the antibiotic over night to select the right strain.
 ===='''2 - ObtaµLining RBS_lacZ_ter. '''====