Template:Team:KULeuven/18 September 2009/BlueLightReceptor

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(Difference between revisions)
(New page: # gel electrophoresis of the RD of ligY and ligA. all of the inserts were ok and ligA had a right signal. # a motherplate for ligY was made # likA and LogA ligations were electroporated)
Line 2: Line 2:
# a motherplate for ligY was made
# a motherplate for ligY was made
# likA and LogA ligations were electroporated
# likA and LogA ligations were electroporated
 +
# a new ligation of likA and logA were made and put overnight.
 +
 +
==saterday==
 +
 +
# enting of the electroporated logA from 18/09. the enting was done on tc(5) as wel as on ap plates. there should not be any growth on the ap plates since the resistence gene is cut in half by the insert. the electroporation of likA didn't show any growth
 +
# a new RD of ligA, pSB3K3 and pBR322 with EcoRI and PstI were done and put overnight.
 +
# an electroporation of likA and logA (ligations from 18/09) was done
 +
 +
==sunday==
 +
 +
# the elecroporation from saterday was not succesfull for logA but there was a colony on the likA plates. this was reented and a liquid culture was made to miniprepp and check.
 +
# the restriction digest from saterday was put on gel and checked. all but one lane had good signal. a gel extraction was done with following results: ligA: 4,8ng/µl ; pBR322: 12,3ng/µl and pSB3K3: 14,3ng/µl (allw with good 260/280 values). pSB3K3 and ligA were used to perform a new ligation (likA)
 +
# the overenting of logA on tc/ap plates from saterday was succesfull. the cultures only grew on the tc plates.  the liquid cultures that were made from these logA's were miniprepped with following results: logA(1): 59,1ng/µl and logA(2): 91,5ng/µl. a restriction digest with EcoRI and PstI was done and put on gel. the signal however showed that only one of the enzymes cut properly. the signal was however on the right height. a new restriction digest will be performed on monday.
 +
# a liquid culture of ligY mother plate was made for glycerol sotck.

Revision as of 09:37, 21 September 2009

  1. gel electrophoresis of the RD of ligY and ligA. all of the inserts were ok and ligA had a right signal.
  2. a motherplate for ligY was made
  3. likA and LogA ligations were electroporated
  4. a new ligation of likA and logA were made and put overnight.

saterday

  1. enting of the electroporated logA from 18/09. the enting was done on tc(5) as wel as on ap plates. there should not be any growth on the ap plates since the resistence gene is cut in half by the insert. the electroporation of likA didn't show any growth
  2. a new RD of ligA, pSB3K3 and pBR322 with EcoRI and PstI were done and put overnight.
  3. an electroporation of likA and logA (ligations from 18/09) was done

sunday

  1. the elecroporation from saterday was not succesfull for logA but there was a colony on the likA plates. this was reented and a liquid culture was made to miniprepp and check.
  2. the restriction digest from saterday was put on gel and checked. all but one lane had good signal. a gel extraction was done with following results: ligA: 4,8ng/µl ; pBR322: 12,3ng/µl and pSB3K3: 14,3ng/µl (allw with good 260/280 values). pSB3K3 and ligA were used to perform a new ligation (likA)
  3. the overenting of logA on tc/ap plates from saterday was succesfull. the cultures only grew on the tc plates. the liquid cultures that were made from these logA's were miniprepped with following results: logA(1): 59,1ng/µl and logA(2): 91,5ng/µl. a restriction digest with EcoRI and PstI was done and put on gel. the signal however showed that only one of the enzymes cut properly. the signal was however on the right height. a new restriction digest will be performed on monday.
  4. a liquid culture of ligY mother plate was made for glycerol sotck.
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