Team:Wageningen UR/Journal/week14

From 2012.igem.org

(Difference between revisions)
(→week 14: 30 july - 5 august)
(→Lab work)
Line 30: Line 30:
'''TuYV'''
'''TuYV'''
-
* The four 'brickable' TuYV CP gene amplicons were digested, ligated into the backbone from BBa_J04450 and transformed into our electrocompetent Mach1 ''E coli''. Despite negative results on two PCR checks, minipreps were done anyway and a third PCR check showed presence of all of our inserts in their respective minipreps!
+
* The four 'brickable' TuYV CP gene amplicons (1,1H,4,4H) were digested, ligated into the backbone from BBa_J04450, which is AMP resistant and transformed into our electrocompetent Mach1 ''E coli'' Mach 1.  
 +
* Colony check: there were many colonies on the sample plate. No colonies were on the negative plate.
 +
* We did two colonies PCR check for the samples, but we got negative results from the colony PCR, but we continued to miniprep those colonoes which were used for colony PCR and check them with normal PCR, this time we saw expected bands.
 +
 
 +
 
 +
 
'''PLRV'''
'''PLRV'''

Revision as of 10:09, 18 September 2012

week 14: 30 july - 5 august

Lab work

Hepatitis B

30.July

  • Miniprepping of successful transformants from the transformation (Hep B + BBa_J04500)
  • Digestion check of those minipreps


1.August

-> the digestion check was repeated 2 times (once with a longer incubation time and once with a higher amount of DNA in the digestion mixture) until the insert size could be seen on the gel


Digestion check 1.August.png

digestion check 1.August

-> the digestion check confirmed the right insert sizes in the samples they where expected (around 800bp)


TuYV

  • The four 'brickable' TuYV CP gene amplicons (1,1H,4,4H) were digested, ligated into the backbone from BBa_J04450, which is AMP resistant and transformed into our electrocompetent Mach1 E coli Mach 1.
  • Colony check: there were many colonies on the sample plate. No colonies were on the negative plate.
  • We did two colonies PCR check for the samples, but we got negative results from the colony PCR, but we continued to miniprep those colonoes which were used for colony PCR and check them with normal PCR, this time we saw expected bands.



PLRV

The PCR products obtained after Reverse Transcription followed by PCR were used as template to add iGEM prefix and suffix.


[previous week] [next week]

Retrieved from "http://2012.igem.org/Team:Wageningen_UR/Journal/week14"