Template:Team:Uppsala/JS/notebook

Revision as of 20:39, 27 September 2013 (view source)

MikaelS (Talk | contribs)

← Older edit

Revision as of 21:20, 27 September 2013 (view source)

MikaelS (Talk | contribs)

Newer edit →

Line 399:

}

-

+

else if(id == 'd2013626')

+

{

+

ds = '<div id="dairy-text"><h2>Wednesday 2013-06-26</h2> Name of participants: Christoffer A, Stephanie H, Alona N, Anders E, Mikael S, Anton B, Viktor B, Marcus H, Lovisa P, Emil M, Ken B-A, Karl H, Theodor L, Magnus B., Malin B., Thorsteinn O., Nils A., Alexander W., Niclas S., Pontus D., Christoffer F. <h2>Ongoing constructs:</h2> L. reuteri: 1. CF48-pLR581 2. 1063-pLUL631 3. DSM 20016-pVs2 4. 100-23-noplasmid 6. DSM 20016-pLUL63TsA8 7. DSM 20016-pGFP 8. 100-23-pGT232 12. DSM 20016-noplasmid 14. DSM 20016-pLUL631(B?) L. plantarum: 5. 256-rifR-pAMβ1 10. 256-noplasmid 11. 36E-noplasmid E. faecalis: 9. JH2-2 pAMβ1 L. lactis 13. MG1363-pJP059 15. unknown-pGus 16. MG1363-noplasmid E.coli: 14. pET13b-zifz:4cl-PBSII:STS 15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT 16. pSB1C3-TAL_m 17. pSB1C3-4Cl_m 18. pSB1C3-CHS_m 25.pSB1C3-CrtY 19.psB4C5-CrtE~O-Z / w. linker, ZF & RBS 20.psB4C5-CrtE~Z / w. linker, ZF & RBS 21.psB4C5-CrtE~I / w. linker, ZF & RBS 42. D5α with plasmid pSB4S15-red <h2>Todays work</h2> Competent cells preparation: Lb11. plantarum 36E no plasmid Lb10. plantarum 256 no plasmid Lb12. reuteri DSM 20016 no plasmid Transformation: plantarum 36E pAMB1 plantarum 256 pAMB1 Lb3. reuteri DSM 20016 pVS2 25.pSB1C3-CrtY 42. pSB4S15-red O/N: Lb5.2 plantarum 256 pAMβ1* Lb8.2 reuteri 100-23 pGT232* Lb13. Lc. lactis MG1363 pJP059** Lb15. Lc. lactis MG1363 pGus** Lb16. Lc. lactis MG1363 no plasmid** *Two tubes of 8.2 were prepared. It has grown rather poorly to not at all before. If it continues to grow poorly we may have to redo O/N from a plate rather than from our frozen stock. ** Optimized conditions → 10µg/ml antibiotics, fill almost up the falcon tube with M17-broth to get rid of the air, let grow in 30o C, make sure that the bacteria pellet is dissolved. Digestion: 19.1.psB4C5-CrtE~O-Z / w. linker, ZF & RBS 19.2psB4C5-CrtE~O-Z / w. linker, ZF & RBS 20.1.psB4C5-CrtE~Z / w. linker, ZF & RBS 20.2.psB4C5-CrtE~Z / w. linker, ZF & RBS 21.1.psB4C5-CrtE~I / w. linker, ZF & RBS 21.2.psB4C5-CrtE~I / w. linker, ZF & RBS Gel electrophoresis: 15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT PCR amplification of construct: 15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT (third try, using same concentration of template as second try but lower annealing temperature ) 19.1.psB4C5-CrtE~O-Z / w. linker, ZF & RBS 19.2psB4C5-CrtE~O-Z / w. linker, ZF & RBS 20.1.psB4C5-CrtE~Z / w. linker, ZF & RBS 20.2.psB4C5-CrtE~Z / w. linker, ZF & RBS 21.1.psB4C5-CrtE~I / w. linker, ZF & RBS 21.2.psB4C5-CrtE~I / w. linker, ZF & RBS <h2>Results</h2> Previous day: O/N: Lb13. Lc. lactis MG1363 pJP059: Growing, visual pellet.* Lb15. Lc. lactis MG1363 pGus: Growing, visual pellet.* Lb16. Lc. lactis MG1363 no plasmid: Growing, visual pellet.* *Weak growth in the morning, better in the afternoon. → Let grow for one more day + new O/N with optimized conditions. Streaking of strains: Lb13. Lc. lactis MG1363 pJP059: No visual growth → Let grow for one more day. Lb15. Lc. lactis MG1363 pGus: No visual growth → --”-- Lb16. Lc. lactis MG1363 no plasmid: No visual growth → --”— Today: Gel electrophoresis: 15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT: Failed, possibly nonspecific primer binding, not enough template DNA. Image 1 (2013.06.26) Transformation: 25.pSB1C3-CrtY: failed D5alpha competent cells were competent <h2>Other experiments</h2> Preparation of cold electroporation buffer for competent cells Preparation of 50% glycerol stock: 500 ml 50% glycerol stock was prepared from 86-88% glycerol.</div>'

+

}

else if(id == 'd201373')

{

@@ Line 399: / Line 399: @@
   }
+else if(id == 'd2013626')
+  {
+	ds = '<div id="dairy-text"><h2>Wednesday 2013-06-26</h2><br><b>Name of participants: </b>Christoffer A, Stephanie H, Alona N, Anders E, Mikael S, Anton B, Viktor B, Marcus H, Lovisa P, Emil M, Ken B-A, Karl H, Theodor L, Magnus B., Malin B., Thorsteinn O., Nils A., Alexander W., Niclas S., Pontus D., Christoffer F.<br><br><h2>Ongoing constructs:</h2><br><b>L. reuteri: </b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>12. DSM 20016-noplasmid<br>14. DSM 20016-pLUL631(B?)<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br>10. 256-noplasmid<br>11. 36E-noplasmid<br><br><b>E. faecalis:</b><br>9. JH2-2 pAMβ1<br><br><b>L. lactis</b><br>13. MG1363-pJP059<br>15. unknown-pGus<br>16. MG1363-noplasmid<br><br><b>E.coli:</b><br>14. pET13b-zifz:4cl-PBSII:STS<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br>25.pSB1C3-CrtY<br>19.psB4C5-CrtE~O-Z / w. linker, ZF & RBS<br>20.psB4C5-CrtE~Z / w. linker, ZF & RBS<br>21.psB4C5-CrtE~I / w. linker, ZF & RBS<br>42. D5α with plasmid pSB4S15-red<br><br><h2>Todays work</h2><br><b>Competent cells preparation:</b><br>Lb11. plantarum 36E no plasmid<br>Lb10. plantarum 256 no plasmid<br>Lb12. reuteri DSM 20016 no plasmid<br><br><b>Transformation:</b><br>plantarum 36E pAMB1<br>plantarum 256 pAMB1<br>Lb3. reuteri DSM 20016 pVS2<br>25.pSB1C3-CrtY<br>42. pSB4S15-red <br><br><b>O/N:</b><br>Lb5.2 plantarum 256 pAMβ1*<br>Lb8.2 reuteri 100-23 pGT232*<br>Lb13. Lc. lactis MG1363 pJP059**<br>Lb15. Lc. lactis MG1363 pGus**<br>Lb16. Lc. lactis MG1363 no plasmid**<br>*Two tubes of 8.2 were prepared. It has grown rather poorly to not at all before. If it continues to grow poorly we may have to redo O/N from a plate rather than from our frozen stock.<br>** Optimized conditions → 10µg/ml antibiotics, fill almost up the falcon tube with M17-broth to get rid of the air, let grow in 30o C, make sure that the bacteria pellet is dissolved.  <br><br><b>Digestion:</b><br>19.1.psB4C5-CrtE~O-Z / w. linker, ZF & RBS<br>19.2psB4C5-CrtE~O-Z / w. linker, ZF & RBS<br>20.1.psB4C5-CrtE~Z / w. linker, ZF & RBS<br>20.2.psB4C5-CrtE~Z / w. linker, ZF & RBS<br>21.1.psB4C5-CrtE~I / w. linker, ZF & RBS<br>21.2.psB4C5-CrtE~I / w. linker, ZF & RBS<br><br><b>Gel electrophoresis:</b><br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br><br><b>PCR amplification of construct:</b><br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT (third try, using same concentration of template as second try but lower annealing temperature )<br>19.1.psB4C5-CrtE~O-Z / w. linker, ZF & RBS<br>19.2psB4C5-CrtE~O-Z / w. linker, ZF & RBS<br>20.1.psB4C5-CrtE~Z / w. linker, ZF & RBS<br>20.2.psB4C5-CrtE~Z / w. linker, ZF & RBS<br>21.1.psB4C5-CrtE~I / w. linker, ZF & RBS<br>21.2.psB4C5-CrtE~I / w. linker, ZF & RBS<br><br><br><h2>Results</h2><br><b>Previous day:</b><br><b>O/N:</b> <br>Lb13. Lc. lactis MG1363 pJP059: Growing, visual pellet.*<br>Lb15. Lc. lactis MG1363 pGus: Growing, visual pellet.*<br>Lb16. Lc. lactis MG1363 no plasmid: Growing, visual pellet.*<br>*Weak growth in the morning, better in the afternoon. → Let grow for one more day + new O/N with optimized conditions.<br><br><b>Streaking of strains: </b><br>Lb13. Lc. lactis MG1363 pJP059: No visual growth → Let grow for one more day.<br>Lb15. Lc. lactis MG1363 pGus: No visual growth →		--”--<br>Lb16. Lc. lactis MG1363 no plasmid: No visual growth → 	--”—<br><br><b>Today:</b><br><b>Gel electrophoresis:</b><br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT: Failed, possibly nonspecific primer binding, not enough template DNA. Image 1 (2013.06.26)<br><br><b>Transformation:</b><br>25.pSB1C3-CrtY: failed<br>D5alpha competent cells were competent<br><br><br><h2>Other experiments</h2><br><b>Preparation of cold electroporation buffer for competent cells</b><br><br><b>Preparation of 50% glycerol stock: </b><br>500 ml 50% glycerol stock was prepared from 86-88% glycerol.</div>'
+ }
   else if(id == 'd201373')
   {

Template:Team:Uppsala/JS/notebook

From 2013.igem.org

Revision as of 21:20, 27 September 2013