Team:Hong Kong HKUST/wetlab/notebook

From 2014.igem.org

(Difference between revisions)
Line 834: Line 834:
·      Preparation for submittion of samples to be sequenced<br>
·      Preparation for submittion of samples to be sequenced<br>
·      Inoculation of pSB1C3-Pcombox (100bp, 150bp, 160bp, 180bp and 249bp)<br>
·      Inoculation of pSB1C3-Pcombox (100bp, 150bp, 160bp, 180bp and 249bp)<br>
-
·      <br>
+
·      colony PCR for ligation product part 1<br>
-
·      <br>
+
·      colony PCR for ligation product part 2<br>
-
·      <br>
+
·      colony PCR for gibson assembly<br>
-
·      <br>
+
·      Run gel for PCR products<br>
-
·      <br>
+
·      comE-D58E plasmid extraction<br>
·   <br>
·   <br>
Line 862: Line 862:
·      Gel purification of digested pSB1C3-Pcombox (150bp, 160bp, 180bp and 249bp)<br>
·      Gel purification of digested pSB1C3-Pcombox (150bp, 160bp, 180bp and 249bp)<br>
·   Inoculation of pSB1C3-BBa_E0240<br>
·   Inoculation of pSB1C3-BBa_E0240<br>
 +
·      Gel extraction and purification of E0240<br>
 +
·      Bacterial inoculation of PcomX (PcomCDE(3))<br>
 +
·      Gel extraction and purification of comE mutant<br>
 +
·      <br>
 +
·      <br>
·      <br>
·      <br>
·   <br>
·   <br>
Line 876: Line 881:
·      Transformation of pSB1C3-Pcombox-BBa_E0240 (160bp and 249bp)<br>
·      Transformation of pSB1C3-Pcombox-BBa_E0240 (160bp and 249bp)<br>
·      Digestion of pSB1C3-BBa_E0240 at XbaI and PstI<br>
·      Digestion of pSB1C3-BBa_E0240 at XbaI and PstI<br>
-
·      <br>
+
·      Gel purification of E0240 insert<br>
-
·      <br>
+
·      Digest PcomX(PcomCDE(3))-pSB1C3 w/ SpeI and PstI<br>
-
·      <br>
+
·      Make aliquotes for oligo - PcomCDE<br>
 +
·      PcomCDE Oligo annealing<br>
 +
·      Ligation of comE mutant insert with K880005<br>
 +
·      Ligation of comE insert with K880005<br>
 +
·      Gel electrophoresis of Gibson assembly mastermix checking<br>
 +
·      Plasmid extraction of J04450(CHL), PcomX(PcomCDE(3)) <br>
 +
·      Bacterial inoculation of comE WT, J04450 (CHL)<br>
·   <br>
·   <br>
<br><b>Aug 20</b> <br>
<br><b>Aug 20</b> <br>
Line 894: Line 905:
·      Colony PCR for ligated pSB1C3-Pcombox-BBa_E0240 (160bp and 249bp)<br>
·      Colony PCR for ligated pSB1C3-Pcombox-BBa_E0240 (160bp and 249bp)<br>
·   Inoculation of  pSB1C3-Pcombox-BBa_E0240 (160bp and 249bp)<br>
·   Inoculation of  pSB1C3-Pcombox-BBa_E0240 (160bp and 249bp)<br>
-
·      <br>
+
·      Ligation of E0240 insert with PcomX<br>
-
·      <br>
+
·      Transformation [E0240-PcomCDE exp+ -ve, pSB1C3-PcomCDE exp+ -ve]<br>
 +
·      Ligation of comED58E insert with K880005<br>
 +
·      Gel extract comE WT<br>
 +
·      PCR check for comED58E-myc<br>
 +
·      Digest J04450(CHL) w/ EcoRI, SpeI, Gel extraction<br>
 +
·      Plasmid extraction of comE WT, J04450 (CHL)<br>
·   <br>
·   <br>
<br><b>Aug 21</b> <br>
<br><b>Aug 21</b> <br>
Line 911: Line 927:
·      Inoculation of pSB1C3-Pcombox-BBa_E0240 (100bp, 150bp and 180bp)<br>
·      Inoculation of pSB1C3-Pcombox-BBa_E0240 (100bp, 150bp and 180bp)<br>
·   Received sequencing result of pSB1C3-Pcombox (100bp, 150bp and 180bp)<br>
·   Received sequencing result of pSB1C3-Pcombox (100bp, 150bp and 180bp)<br>
-
·      <br>
+
·      Ligate PcomX with E0240 insert<br>
-
·   <br>
+
·     PCR, Restrict check Part I (PcomCDE)<br>
-
·      <br>
+
·      O/N Ligation w/ PcomCDE<br>
 +
·      Extract comE WT-pSB1C3<br>
 +
·      PCR and restrict check comED58E-myc<br>
 +
·      Glycerol stock for PcomX <br>
·   <br>
·   <br>
<br><b>Aug 22</b> <br>
<br><b>Aug 22</b> <br>
Line 923: Line 942:
·      PCR of pSB1C3-Pcombox-BBa_E0240 (160bp and 249bp) using Vent polymerase <br>
·      PCR of pSB1C3-Pcombox-BBa_E0240 (160bp and 249bp) using Vent polymerase <br>
·      Sent out pSB1C3-Pcombox-BBa_E0240 (160bp and 249bp) for commercial sequencing<br>
·      Sent out pSB1C3-Pcombox-BBa_E0240 (160bp and 249bp) for commercial sequencing<br>
-
·      <br>
+
·      Transformation of newly ligated (E0240)<br>
-
·      <br>
+
·      Bacterial inoculation of checked colony (E0240)<br>
-
·      <br>
+
·      O/N ligation w/ pSB1C3<br>
-
·      <br>
+
·      Ligation comE WT-pSB1C3<br>
-
·      <br>
+
·      Ligate comED58E-myc w/ K880005<br>
·   <br>
·   <br>
Line 948: Line 967:
·      Digestion of pSB1C3-Pcombox (100bp, 150bp and 160bp, 180bp and 249bp) at XbaI and PstI<br>
·      Digestion of pSB1C3-Pcombox (100bp, 150bp and 160bp, 180bp and 249bp) at XbaI and PstI<br>
·      Gel purification of digested Pcombox-BBa_E0240 (100bp, 150bp and 180bp) insert<br>
·      Gel purification of digested Pcombox-BBa_E0240 (100bp, 150bp and 180bp) insert<br>
 +
·      Ligation of PcomX-E0240<br>
 +
·      Digestion of PcomCDE-E0240<br>
 +
·      Colony PCR pSB1C3-PcomCDE, K880005-comE WT, K880005-comED58E-myc<br>
·      <br>
·      <br>
·      <br>
·      <br>
Line 961: Line 983:
·      Transformation of pSB1C3-K880005.comW-FLAG<br>
·      Transformation of pSB1C3-K880005.comW-FLAG<br>
·      Ligation of comX, 100bp Pcombox-BBa_E0240 and pSB3K3 backbone<br>
·      Ligation of comX, 100bp Pcombox-BBa_E0240 and pSB3K3 backbone<br>
 +
·      Gel pSB1C3-PcomCDE<br>
 +
·      PcomX-E0240 Colony PCR check<br>
 +
·      PcomCDE glycerol stock <br>
 +
·      Restrict check K880005-comE WT<br>
 +
·      Ligation w/ comE WT<br>
 +
·      Restrict check K880005-comED58E-myc<br>
 +
·      <br>
 +
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
Line 975: Line 1,005:
·      Ligation of comX, 100bp Pcombox-BBa_E0240 and pSB3K3 backbone<br>
·      Ligation of comX, 100bp Pcombox-BBa_E0240 and pSB3K3 backbone<br>
·      Inoculation of pSB1C3-Pcombox-BBa_E0240 (100bp, 180bp and 249bp), pSB1C3-Pcombox (160bp)<br>
·      Inoculation of pSB1C3-Pcombox-BBa_E0240 (100bp, 180bp and 249bp), pSB1C3-Pcombox (160bp)<br>
 +
·      pSB1C3-PcomCDE<br>
 +
·      PcomCDE sequencing <br>
 +
·      PCR check K880005-comE WT<br>
 +
·      PCR check K880005-comED58E-myc<br>
 +
·      Ligate K880005-comE-myc<br>
 +
·      <br>
 +
·      <br>
 +
·      <br>
 +
·      <br>
 +
·      <br>
·   <br>
·   <br>
<br><b>Aug 28</b> <br>
<br><b>Aug 28</b> <br>
Line 985: Line 1,025:
·      Restriction check of pSB1C3-Pcombox-BBa_E0240 (180bp and 249bp)<br>
·      Restriction check of pSB1C3-Pcombox-BBa_E0240 (180bp and 249bp)<br>
·      Colony PCR of pSB3K3-comX-100bp Pcombox-BBa_E0240<br>
·      Colony PCR of pSB3K3-comX-100bp Pcombox-BBa_E0240<br>
-
·      <br>
+
·      pSB1C3-PcomCDE overnight ligation<br>
-
·      <br>
+
·      PcomX-E0240 ligation<br>
 +
·      Gel extract I20260 and E0240<br>
 +
·      Glycerol stock for K880005-comE WT<br>
 +
·      Colony PCR for K880005-comED58E-myc  <br>
 +
·      Ligate K880005-comE-myc <br>
 +
·      PCR comED58E-myc<br>
 +
·      Ligation of Part 1 and Part 2<br>
 +
 
·   <br>
·   <br>
<br><b>Aug 29</b> <br>
<br><b>Aug 29</b> <br>
Line 996: Line 1,043:
·      Colony PCR for pSB3K3-comX-100bp Pcombox-BBa_E0240<br>
·      Colony PCR for pSB3K3-comX-100bp Pcombox-BBa_E0240<br>
·      Ligation of pSB1C3-160bp Pcombox with BBa_E0240 insert<br>
·      Ligation of pSB1C3-160bp Pcombox with BBa_E0240 insert<br>
-
·      <br>
+
·      Transform pSB1C3-PcomCDE <br>
-
·      <br>
+
·      Transform PcomX-E0240 <br>
-
·      <br>
+
·      Ligate I20260 and E0240 <br>
 +
·      Ligate K880005-comE WT  <br>
 +
·      Ligate K880005-comE-myc<br>
 +
·      Ligate K880005-comED58E-myc  <br>
 +
·      Gel check comED58E-myc <br>
·   <br>
·   <br>
Line 1,031: Line 1,082:
·      Inoculation of pSB1C3-K880005.comX-myc.B0015.Combox.GFP<br>
·      Inoculation of pSB1C3-K880005.comX-myc.B0015.Combox.GFP<br>
·      Received the sequencing result of pSB1C3-160bp Pcombox<br>
·      Received the sequencing result of pSB1C3-160bp Pcombox<br>
-
·      <br>
+
·      Colony PCR: pSB1C3-PcomCDE, PcomX-E0240, I20260+Part2, E0240+Part2, E0240+Part2, K880005-comE WT, K990005-comE-myc, K990005-comEd58E-myc<br>
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
·   <br>
·   <br>
Line 1,043: Line 1,090:
·      Re-digestion of pSB1C3-K880005.comX-myc.B0015.Combox.GFP on EcoRI-HF and PstI-HF, gel extraction and gel purification<br>
·      Re-digestion of pSB1C3-K880005.comX-myc.B0015.Combox.GFP on EcoRI-HF and PstI-HF, gel extraction and gel purification<br>
·      Restriction check of pSB1C3-160bp Pcombox-BBa_E0240 at XmnI and NcoI<br>
·      Restriction check of pSB1C3-160bp Pcombox-BBa_E0240 at XmnI and NcoI<br>
-
·      <br>
+
·      Whole construct gel check <br>
-
·      <br>
+
·      Restrict check PcomX-E0240<br>
-
·      <br>
+
·      Restrict check Part 1 + Part 2<br>
-
·      <br>
+
·      Restrict check I20260 and E0240 <br>
-
·      <br>
+
·      Restrict check K880005-comE WT<br>
-
·      <br>
+
·      Restrict check K880005-comE-myc<br>
 +
·      Restrict check K880005-comED58E-myc (XbaI & PstI)<br>
·   <br>
·   <br>
Line 1,056: Line 1,104:
·      Inoculation of pSB1C3-K880005.comW-FLAG.B0015<br>
·      Inoculation of pSB1C3-K880005.comW-FLAG.B0015<br>
·      Restriction check of pSB1C3-160bp Pcombox-BBa_E0240 at NcoI and NotI<br>
·      Restriction check of pSB1C3-160bp Pcombox-BBa_E0240 at NcoI and NotI<br>
-
·      <br>
+
·      Restrict check I20260-Part 2 (3 tubes) (XbaI & PstI)<br>
-
·      <br>
+
·      Restrict check E0240-Part 2 (2 tubes)<br>
-
·      <br>
+
·      Restrict check Whole construct (1 tubes)<br>
-
·      <br>
+
·      Restrict check -ve of whole construct ligation (5 tubes) <br>
-
·      <br>
+
·      Restrict check pSB1C3-PcomCDE (3 tubes) (NcoI & PstI)<br>
-
·      <br>
+
·   <br>
·   <br>
Line 1,072: Line 1,119:
·      Digestion of pSB1C3-160bp Pcombox-BBa_E0240 at XbaI and PstI<br>
·      Digestion of pSB1C3-160bp Pcombox-BBa_E0240 at XbaI and PstI<br>
·      Ligation of 160bp Pcombox-BBa_E0240,comX and pSB3K3<br>
·      Ligation of 160bp Pcombox-BBa_E0240,comX and pSB3K3<br>
-
·      <br>
+
·      Restrict check I20260-Part 2, E0240-Part 2, I20260-Part 2, whole construct, -ve of whole construct ligation XP<br>
-
·      <br>
+
·      Restrict check PcomCDE-E0240<br>
-
·      <br>
+
·      Restrict check I20260, E0240, Whole construct 3, 4,5 NcoI lambda ladder<br>
 +
·      Restrict check and colony PCR for pSB1C3-PcomCDE NcoI<br>
 +
·      Restrict check K880005+comED58E-N-myc<br>
 +
·      Restrict check  K880005+comE-C-myc<br>
·   <br>
·   <br>
Line 1,080: Line 1,130:
Wet lab<br>
Wet lab<br>
·      Inoculation of pSB3K3-Combox.GFP<br>
·      Inoculation of pSB3K3-Combox.GFP<br>
-
·      <br>
+
·      Sequencing for pSB1C3-PcomCDE<br>
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
·      <br>
·      <br>
·   <br>
·   <br>
Line 1,099: Line 1,143:
Wet lab<br>
Wet lab<br>
·      Digestion check of pSB3K3-Combox.GFP on XmnI<br>
·      Digestion check of pSB3K3-Combox.GFP on XmnI<br>
-
·      <br>
+
·      Restrict check and plasmid PCR for PcomCDE-E0240<br>
-
·      <br>
+
·      Obtain sequencing results for PcomCDE-pSB1C3<br>
-
·      <br>
+
·      Whole construct (PcomX)<br>
-
·      <br>
+
·      K880005-comE/comED58E-myc digest w/ EcoRI and XbaI<br>
-
·      <br>
+
·      Ligation of Part 1 and 2 (also I20260, E0240) (1200)<br>
-
·      <br>
+
·      Transformation (1400)<br>
-
·      <br>
+
 
-
·      <br>
+
·   <br>
·   <br>
Line 1,112: Line 1,155:
<br><b>Sept 9</b> <br>
<br><b>Sept 9</b> <br>
Wet lab<br>
Wet lab<br>
-
·      <br>
 
-
·      <br>
 
-
·      <br>
 
-
·      <br>
 
-
·      <br>
 
-
·      <br>
 
-
·      <br>
 
-
·      <br>
 
-
·      <br>
 
·   <br>
·   <br>
Line 1,126: Line 1,160:
Wet lab<br>
Wet lab<br>
·      Streak plate for pSB3K3-Combox.GFP<br>
·      Streak plate for pSB3K3-Combox.GFP<br>
-
·      <br>
+
·      Restrict check PcomCDE-E0240<br>
-
·      <br>
+
·      Obtain sequencing results for PcomCDE-pSB1C3<br>
-
·      <br>
+
·      Whole construct (PcomX)<br>
-
·      <br>
+
·      K880005-comE/comED58E-myc send to sequencing<br>
-
·      <br>
+
·      Ligation and transformation of Part 1 and 2 (also I20260, E0240)<br>
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
·   <br>
·   <br>
Line 1,139: Line 1,170:
Wet lab<br>
Wet lab<br>
·      Restriction check of pSB3K3-comX-160bp Pcombox-BBa_E0240 at EcoRI and PstI<br>
·      Restriction check of pSB3K3-comX-160bp Pcombox-BBa_E0240 at EcoRI and PstI<br>
-
·      <br>
+
·      Obtain sequencing results PcomCDE-pSB1C3<br>
-
·      <br>
+
·      Gel electrophoresis of colony PCR product from 10/9 PcomCDE-E0240<br>
-
·      <br>
+
·      Bacterial inoculation of checked products (for sequencing) and streak plates<br>
-
·      <br>
+
·      Check colony fluorescene for whole construct (PcomX)<br>
-
·      <br>
+
·      Gel extraction and purification of E0240 and I20260<br>
-
·      <br>
+
·      Gel extraction and purification of PcomCDE-E0240 (EcoRI and SpeI)<br>
-
·      <br>
+
·      Streak plate for K880005-comE-C-myc<br>
-
·      <br>
+
 
·   <br>
·   <br>
Line 1,153: Line 1,184:
·      Restriction check of pSB3K3-160bp Pcombox-BBa_E0240 at EcoRI and PstI<br>
·      Restriction check of pSB3K3-160bp Pcombox-BBa_E0240 at EcoRI and PstI<br>
·      Transformation of pSB3K3-160bp Pcombox-BBa_E0240<br>
·      Transformation of pSB3K3-160bp Pcombox-BBa_E0240<br>
-
·      <br>
+
·      Make glycerol stock for PcomCDE-pSB1C3<br>
-
·      <br>
+
·      Gel electrophoresis of colony PCR product of PcomCDE-E0240 from 10/9; Bacterial inoculation of checked products (for sequencing); Streak plate<br>
-
·      <br>
+
·      Check colony and fluorescence for whole construct (PcomX)<br>
-
·      <br>
+
·      Streak plate for K880005-comE-C-myc<br>
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
·   <br>
·   <br>
       </div>
       </div>
Line 1,169: Line 1,197:
Wet lab<br>
Wet lab<br>
·      Inoculation of pSB1C3-K880005.comW-FLAG<br>
·      Inoculation of pSB1C3-K880005.comW-FLAG<br>
-
·      <br>
+
·      PCR check and inoculation of colonies of I20260/E0240-K880005-comE-C-myc, I20260/E0240-K880005-comED58E-N-myc<br>
-
·      <br>
+
·      Restrict check, Gel extraction and Gel purification of pSB3K3 J04450 (CHL)<br>
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
·   <br>
·   <br>
Line 1,183: Line 1,205:
Wet lab<br>
Wet lab<br>
·      Digestion of pSB1C3-K880005.comW-FLAG on EcoRI-HF and SpeI<br>
·      Digestion of pSB1C3-K880005.comW-FLAG on EcoRI-HF and SpeI<br>
-
·      <br>
 
-
·      <br>
 
-
·      <br>
 
-
·      <br>
 
-
·      <br>
 
-
·      <br>
 
-
·      <br>
 
-
·      <br>
 
·   <br>
·   <br>
Line 1,201: Line 1,215:
·      Digestion check of Combox + GFP on XmnI and PstI<br>
·      Digestion check of Combox + GFP on XmnI and PstI<br>
·      Restriction check of pSB3K3-160bp Pcombox-BBa_E0240 at NotI and PstI<br>
·      Restriction check of pSB3K3-160bp Pcombox-BBa_E0240 at NotI and PstI<br>
-
·      <br>
+
·      PCR check and inoculation for I20260/E0240-K880005-comE-C-myc, I20260/E0240-K880005-comED58E-N-myc<br>
-
·      <br>
+
·      Restrict check pSB3K3 (already in stock)<br>
-
·      <br>
+
·      Gel extraction and Gel purification of J04450 (CHL) insert X,S<br>
 +
·      Sequencing reaction for K880005-comED58E-N-myc, K880005-comE-C-myc<br>
·   <br>
·   <br>
Line 1,210: Line 1,225:
·      Ligation of comX-myc into pSB1C3 cut on XbaI and PstI<br>
·      Ligation of comX-myc into pSB1C3 cut on XbaI and PstI<br>
·      Inoculation of pSB1C3-K880005.comW-FLAG.B0015<br>
·      Inoculation of pSB1C3-K880005.comW-FLAG.B0015<br>
-
·      <br>
+
·      Colony PCR and inoculation of correct colonies for I20260/E0240-K880005-comE-C-myc, I20260/E0240-K880005-comED58E-N-myc<br>
-
·      <br>
+
·      Gel extraction and gel purification for J04450 (CHL), J04450 (KAN)<br>
-
·      <br>
+
·      Sequencing reaction and Restrict check for K880005-comED58E-N-myc, K880005-comED58E-N-myc<br>
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
·   <br>
·   <br>
Line 1,222: Line 1,233:
Wet lab<br>
Wet lab<br>
·      Digestion check of pSB1C3-K880005.comW-FLAG.B0015 on XbaI and PstI<br>
·      Digestion check of pSB1C3-K880005.comW-FLAG.B0015 on XbaI and PstI<br>
-
·      <br>
+
·      Colony PCR, inoculation for I20260/E0240-K880005-comED58E-N-myc<br>
-
·      <br>
+
·      Miniprep for I20260/E0240-K880005-comE-C-myc<br>
-
·      <br>
+
·      Gel extraction and gel purification J04450 (CHL), J04450 (KAN) XS<br>
-
·      <br>
+
·      Sequencing reaction for K880005-comE-C-myc<br>
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
·   <br>
·   <br>
Line 1,243: Line 1,250:
·      Inoculation of pSB1C3-K880005.comW-FLAG.B0015<br>
·      Inoculation of pSB1C3-K880005.comW-FLAG.B0015<br>
·      Ligation of pSB3K3 backbone with 160bp Pcombox-BBa_E0240<br>
·      Ligation of pSB3K3 backbone with 160bp Pcombox-BBa_E0240<br>
-
·      <br>
+
·      Gel extraction and gel purification of I20260/E0240-K880005-comED58E-N-myc EP<br>
-
·      <br>
+
·      Colony PCR and inoculation of I20260/E0240-K880005-comE-C-myc <br>
-
·      <br>
+
·      Gel extraction and gel purification J04450 (CHL) (X&S), J04450 (KAN) (E&P)  <br>
-
·      <br>
+
·      Oligo annealing and overnight ligation w/ pSB1C3for PcomCDE<br>
-
·      <br>
+
·   <br>
·   <br>
Line 1,256: Line 1,262:
·      Ligation of pSB1C3-K880005.comX into pSB1C3-B0015 cut on EcoRI-HF and XbaI<br>
·      Ligation of pSB1C3-K880005.comX into pSB1C3-B0015 cut on EcoRI-HF and XbaI<br>
·      Transformation of pSB3K3-160bp Pcombox-BBa_E0240 ligated on Sept 22<br>
·      Transformation of pSB3K3-160bp Pcombox-BBa_E0240 ligated on Sept 22<br>
-
·      <br>
+
·      Gel check and redo liagtion for I20260-K880005-comED58E-N-myc, E0240-K880005-comED58E-N-myc, I20260-K880005-comE-C-myc, E0240-K880005-comE-C-myc<br>
-
·      <br>
+
·      Gel extraction and purification for I20260-K880005-comED58E-N-myc, E0240-K880005-comED58E-N-myc<br>
-
·      <br>
+
·      Ethanol precipitation for sequencing, restrict check  K880005-comED58E-N-myc, K880005-comE-C-myc<br>
-
·      <br>
+
·      Dephosphorylation and gel extraction and gel purification of J04450 (CHL) (X&S)<br>
-
·      <br>
+
·      Gel extraction and Gel purification, dephosphorylation of J04450 (KAN) (E&P)<br>
 +
·      Oligo annealing, overnight ligation w/ pSB1C3 of PcomCDE<br>
·   <br>
·   <br>
Line 1,267: Line 1,274:
·      Digestion check of pSB1C3-K880005.comX.B0015 on XbaI and PstI, gel extraction and gel purification<br>
·      Digestion check of pSB1C3-K880005.comX.B0015 on XbaI and PstI, gel extraction and gel purification<br>
·      Inoculation of bacterial colonies carrying pSB3K3-160bp Pcombox-BBa_E0240<br>
·      Inoculation of bacterial colonies carrying pSB3K3-160bp Pcombox-BBa_E0240<br>
-
·      <br>
+
·      Restrict check, gel extraction and purification, Ligation w/ pSB3K3 I20260-K880005-comED58E-N-myc, E0240-K880005-comED58E-N-myc, I20260-K880005-comE-C-·      myc, E0240-K880005-comE-C-myc <br>
-
·      <br>
+
·      Obtain sequencing results for K880005-comED58E-N-myc, K880005-comE-C-myc<br>
-
·      <br>
+
·      Transform pSB1C3-PcomCDE<br>
-
·      <br>
+
 
-
·      <br>
+
-
·      <br>
+
-
·      <br>
+
·   <br>
·   <br>

Revision as of 17:08, 16 October 2014



Notebook

Throughout the course of our project, we have continuously recorded our daily work, such as wetlab activities and research findings. The notebook for Pneuumonsensor and Riboregulator are displayed separately.






Pneumosensor

June 2014

Week 1
Week 2
Week 3
Week 4
Week 5

July 2014

Week 1
Week 2
Week 3
Week 4
Week 5

August 2014

Week 1
Week 2
Week 3
Week 4
Week 5

September 2014

Week 1
Week 2
Week 3
Week 4
Week 5





Riboregulator

June 2014

Week 1
Week 2
Week 3
Week 4
Week 5

July 2014

Week 1
Week 2
Week 3
Week 4
Week 5

August 2014

Week 1
Week 2
Week 3
Week 4
Week 5

September 2014

Week 1
Week 2
Week 3
Week 4
Week 5


TOP

Home

Pneumosensor

Riboregulator

Human Practice

Team

WetLab

Achievement