IGEM:MIT/2006/Notebook/2006-10-8

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Current revision (18:27, 8 October 2006) (view source)
 
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-- ligated 320 to OS-Q-119 in an A/C backbone & transformed into both Barry's top10 cells and the IK cells Bo put in the -80 yesterday '''done, VV'''
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#ligated 320 to OS-Q-119 in an A/C backbone & transformed into both Barry's top10 cells and the IK cells Bo put in the -80 yesterday '''done, VV'''
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-- ligated 250 to (250+400) in an A/T backbone & transformed into top10 & IK '''done, VV'''
+
#ligated 250 to (250+400) in an A/T backbone & transformed into top10 & IK '''done, VV'''
also, after talking to Jason, we decided that we want to have 400 on its own, so:
also, after talking to Jason, we decided that we want to have 400 on its own, so:
--transformed the  miniprepped mix of 400  in kan backbone and 250 in A/C backbone into Top10, and plated on kan plates.  so, the transformants we get should be either double transformants (both 400 and 250), or just single 400 transformants.  We want to isolate the 400 (IAGD) so we can ligate it to 250. so, what we need to do is pick the colonies we get tomorrow, and put them in some water...and then take a little out of each tube to plate on an a/c plate.  if cells grow on the a/c plate (it'll take about 3 hrs to tell), then we'll know that they were double tranformants...that is, we don't want them.  then we'll just LC the cells that we know have ONLY 400 in them, and we can miniprep that on tuesday.
--transformed the  miniprepped mix of 400  in kan backbone and 250 in A/C backbone into Top10, and plated on kan plates.  so, the transformants we get should be either double transformants (both 400 and 250), or just single 400 transformants.  We want to isolate the 400 (IAGD) so we can ligate it to 250. so, what we need to do is pick the colonies we get tomorrow, and put them in some water...and then take a little out of each tube to plate on an a/c plate.  if cells grow on the a/c plate (it'll take about 3 hrs to tell), then we'll know that they were double tranformants...that is, we don't want them.  then we'll just LC the cells that we know have ONLY 400 in them, and we can miniprep that on tuesday.

Current revision

  1. ligated 320 to OS-Q-119 in an A/C backbone & transformed into both Barry's top10 cells and the IK cells Bo put in the -80 yesterday done, VV
  2. ligated 250 to (250+400) in an A/T backbone & transformed into top10 & IK done, VV

also, after talking to Jason, we decided that we want to have 400 on its own, so: --transformed the miniprepped mix of 400 in kan backbone and 250 in A/C backbone into Top10, and plated on kan plates. so, the transformants we get should be either double transformants (both 400 and 250), or just single 400 transformants. We want to isolate the 400 (IAGD) so we can ligate it to 250. so, what we need to do is pick the colonies we get tomorrow, and put them in some water...and then take a little out of each tube to plate on an a/c plate. if cells grow on the a/c plate (it'll take about 3 hrs to tell), then we'll know that they were double tranformants...that is, we don't want them. then we'll just LC the cells that we know have ONLY 400 in them, and we can miniprep that on tuesday.

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