|
|
| Line 8: |
Line 8: |
| | *[[Team:Chiba/jk/γ/week 6|week 6 (21,September~27,September)]] | | *[[Team:Chiba/jk/γ/week 6|week 6 (21,September~27,September)]] |
| | | | |
| - |
| |
| - | ----
| |
| - | '''24,August 2008'''
| |
| - | <br>TRANSFORMATION
| |
| - | <br>BBa_J63001(yeast optimized YFP)-->colony
| |
| - | <br>Digestion
| |
| - | <br>BBa_F2620(1)①
| |
| - | <br>BBa_F2620(2)②
| |
| - | <br>Menbren Venus YFP③
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①</td><td>②</td><td>③</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA tamplate</td>
| |
| - | <td>3</td><td>3</td><td>3</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Buffer3</td>
| |
| - | <td>1</td><td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>6</td><td>6</td><td>6</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>EcoRⅠ</td>
| |
| - | <td>0.5</td><td>0.5</td><td>0.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>10</td><td>10</td><td>10</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>->37℃ 30min
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①(afetr digestion)</td><td>②(after digestion)</td><td>③(afetr digestion)</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA tamplate</td>
| |
| - | <td>10</td><td>10</td><td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>loading Dye</td>
| |
| - | <td>2</td><td>2</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>12</td><td>12</td><td>12</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①</td><td>②</td><td>③</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA tamplate</td>
| |
| - | <td>2</td><td>2</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>loading Dye</td>
| |
| - | <td>1</td><td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>3</td><td>3</td><td>3</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>6</td><td>6</td><td>6</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>->135V 30min
| |
| - |
| |
| - | ----
| |
| - | '''25,August,2008'''
| |
| - | <br>PCR
| |
| - | <br>BBa_E2030①
| |
| - | <br>BBa_J33202②
| |
| - | <br>BL21③
| |
| - | <br>BBa_J52008④
| |
| - | <br>BBa_I712019⑤
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA tamplate</td>
| |
| - | <td>1</td><td>1</td><td>1</td><td>1</td>><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>FW primer</td>
| |
| - | <td>5(Venus_YFP_fwd)</td><td>5(LacZ_fwd)</td><td>5(LacZ_fwd)</td><td>5(rLUC_fwd)</td><td>5(fLuc_fwd)</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>VR primer</td>
| |
| - | <td>5(VR)</td><td>5(LacZ_rev)</td><td>5(LacZ_rev)</td><td>5(VR)</td><td>5(VR)</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dNTPmix</td>
| |
| - | <td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>thermo pol buffer</td>
| |
| - | <td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA pol(VENT)</td>
| |
| - | <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>68</td><td>68</td><td>68</td><td>68</td><td>68</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>100</td><td>100</td><td>100</td><td>100</td><td>100</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>->Gel
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA tamplate</td>
| |
| - | <td>1</td><td>1</td><td>1</td><td>1</td>><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>loading Dye</td>
| |
| - | <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>4</td><td>4</td><td>4</td><td>4</td><td>4</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>6</td><td>6</td><td>6</td><td>6</td><td>6</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <br>MINI prep
| |
| - | <br>Venus
| |
| - | <br>BBa_F2620
| |
| - | <br>TRANSFORMATRION
| |
| - | <br>Venus
| |
| - | <br>PUC19
| |
| - | <br>pGFPUV
| |
| - | <br>pGEX Venus YFP
| |
| - | <br>Membren Venus YFP
| |
| - | <br>PCR
| |
| - | <br>BBa_J63001①
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA tamplate</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>FW primer(Venus)</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>VR primer</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dNTPmix</td>
| |
| - | <td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>thermo pol buffer</td>
| |
| - | <td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA pol(VENT)</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>68</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>100</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | ----
| |
| - | '''26,August,2008'''
| |
| - | <br>Gel check
| |
| - | <br>BBa_F2620
| |
| - | <br>BBa_F2620
| |
| - | <br>BBa_J63001
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA tamplate</td>
| |
| - | <td>1</td><td>1</td><td>1</td><td>1</td>><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye</td>
| |
| - | <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>4</td><td>4</td><td>4</td><td>4</td><td>4</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>6</td><td>6</td><td>6</td><td>6</td><td>6</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <br>Digestion
| |
| - | <br>vector:BBa_F2620
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>BBa_F2620</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA tamplate</td>
| |
| - | <td>100</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>SpeⅠ</td>
| |
| - | <td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>PstⅠ</td>
| |
| - | <td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA(x10)</td>
| |
| - | <td>13</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Buffer2</td>
| |
| - | <td>13</td>
| |
| - | </tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>130</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>insert:BBa_J52008,BBa_E2030
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>J52008</td><td>E2030</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA tamplate</td>
| |
| - | <td>30</td><td>30</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>XbaⅠ</td>
| |
| - | <td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>PstⅠ</td>
| |
| - | <td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DpnⅠ</td>
| |
| - | <td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Buffer3</td>
| |
| - | <td>4</td><td>4</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA(x10)</td>
| |
| - | <td>4</td><td>4</td>
| |
| - | </tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>40</td><td>40</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>->37℃ 3hour
| |
| - | <br>->SAP
| |
| - | <br>->37℃ 30min
| |
| - | <br>->65℃ 15min
| |
| - | <br>->Zymo Clean
| |
| - | <br>Ligation
| |
| - | <br>vector:BBa_F2620 insert:BBa_J52008①
| |
| - | <br>vector:BBa_F2620 insert:BBa_E2030②
| |
| - | <br>Negative control:BBa_F2620③
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①</td><td>②</td><td>③</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>vector</td>
| |
| - | <td>5</td><td>5</td><td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>inser</td>
| |
| - | <td>10</td><td>7.5</td><td>0</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Ligase Buffer</td>
| |
| - | <td>4</td><td>4</td><td>4</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Ligase</td>
| |
| - | <td>1</td><td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>0</td><td>2.5</td><td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>20</td><td>20</td><td>20</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>->RT 2hour
| |
| - | <br>->TRANSFORMATION XL10G
| |
| - | <br>->37℃ over night
| |
| - | <br>->colony PCR
| |
| - | <br>->MINI PREP
| |
| - |
| |
| - | ----
| |
| - | '''28,August,2008'''
| |
| - | <br>TIME RESPONCE(liquid)
| |
| - |
| |
| - | ----
| |
| - | '''29,August,2008'''
| |
| - | <br>TIME RESPONCE(solid)
| |
| - |
| |
| - | ----
| |
| - | '''30,August,2008'''
| |
| - | <br>TRANSFORMATION
| |
| - | <br>mcherry
| |
| - | <br>PCR
| |
| - | <br>BBa_J52008(rLuc)
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>BBa_J52008</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA tamplate</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>rLuc_fwd</td>
| |
| - | <td>2.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>VR</td>
| |
| - | <td>2.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>thermo pol Buffer</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dNTP</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Vent pol</td>
| |
| - | <td>0.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>34</td>
| |
| - | </tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>50</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | ----
| |
| - | '''31,August,2008'''
| |
| - | <br>digestion
| |
| - | <br>BBa_R0010①
| |
| - | <br>PCR product(BBa_J52008)②
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①</td><td>②</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA </td>
| |
| - | <td>10</td><td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>SpeⅠ</td>
| |
| - | <td>1</td><td>0</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>PstⅠ</td>
| |
| - | <td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>XbaⅠ</td>
| |
| - | <td>0</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA(x100)</td>
| |
| - | <td>0.5</td><td>0.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Buffer2</td>
| |
| - | <td>5</td><td>0</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Buffer3</td>
| |
| - | <td>0</td><td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH20</td>
| |
| - | <td>28.5</td><td>28.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>50</td><td>50</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>->37℃ 3hour
| |
| - | <br>->①20μl refine
| |
| - | <br>->②20μl refine
| |
| - |
| |
| - | ----
| |
| - | '''1,September,2008'''
| |
| - | <br>Ligation
| |
| - | <br>vector:BBa_R0010 insert:BBa_J52008①
| |
| - | <br>negative control:BBa_R0010②
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①</td><td>②</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>vector</td>
| |
| - | <td>2</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>insert</td>
| |
| - | <td>3</td><td>0</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Ligase Buffer</td>
| |
| - | <td>2</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Ligase</td>
| |
| - | <td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>3</td><td>5</td>
| |
| - | </tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>10</td><td>10</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>->R/T 2hour
| |
| - |
| |
| - | <br>TRANSFORMATION
| |
| - | <br>BBa_R0079(Las promoter)
| |
| - | <br>BBa_R0071(RhlR promoter)
| |
| - | <br>BBa_R0077(cinR promoter+RBS)
| |
| - | <br>BBa_R0078(cnR promoter)
| |
| - | <br>BBa_R0062(LUX promoter)
| |
| - |
| |
| - |
| |
| - | ----
| |
| - | '''2,September,2008'''
| |
| - | <br>TIME RESPONCE (Solid)
| |
| - |
| |
| - | <br>COLONY PCR
| |
| - | <br>BBa_R0010+BBa_J52008
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>culture</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <td>Fwd primer</td>
| |
| - | <td>1.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Rev primer</td>
| |
| - | <td>1.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Thermo pol Buffer</td>
| |
| - | <td>3</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dNTP mix</td>
| |
| - | <td>3</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Taq DNA pol (NEB)</td>
| |
| - | <td>0.2 (1 unit)</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>19.8</td>
| |
| - | </tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>30</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>MINI Prep
| |
| - | <br>BBa_R0079
| |
| - | <br>BBa_R0071
| |
| - | <br>BBa_R0077
| |
| - | <br>BBa_R0078
| |
| - | <br>BBa_R0062
| |
| - |
| |
| - | ----
| |
| - | '''3,September,2008'''
| |
| - | <br>Digestion test
| |
| - | <br>BBa_R0079①
| |
| - | <br>BBa_R0071②
| |
| - | <br>BBa_R0077③
| |
| - | <br>BBa_R0078④
| |
| - | <br>BBa_R0062⑤
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①~⑤</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA tamplate</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Buffer3</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>4</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>EcoRⅠ</td>
| |
| - | <td>0.1</td>
| |
| - | </tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>10</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>->37℃30min
| |
| - | <br>->Gel cheack
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①~⑤</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA(digestion sample)</td>
| |
| - | <td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>loading Dye</td>
| |
| - | <td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>12</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | ----
| |
| - | '''4,Septempber,2008'''
| |
| - |
| |
| - |
| |
| - |
| |
| - | ----
| |
| - | '''5,September,2008'''
| |
| - | <br>TIME RESPONCE(liquid)
| |
| - | <br>Digestion test
| |
| - | <br>BBa_J52008+BBa_F2620
| |
| - | <br>BBa_E2030+BBa_F2620
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>each</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA(digestion sample)</td>
| |
| - | <td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>loading Dye</td>
| |
| - | <td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>12</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <br>Luciferase cheack->no light
| |
| - |
| |
| - | ----
| |
| - | '''7,September,2008'''
| |
| - | <br>Cloning
| |
| - | <br>Digestion
| |
| - | <br>vector:BBa_R0040(ptet)① 75ng/μl
| |
| - | <br>insert:BBa_S03158(RBS+RhlR)② 33.6ng/μl
| |
| - | <br>insert:BBa_S03156(RBS+lasR)③ 33.6ng/μl
| |
| - | <br>insert:BBa_S03160(RBS+cinR)④ 38.4ng/μl
| |
| - | <br>vector:BBa_R0010(pLac)⑤ 100ng/μl
| |
| - | <br>insert:BBa_J52008(RBS+rLuc)⑥ 33.6ng/μl
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td><td>⑥</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA(digestion sample)</td>
| |
| - | <td>50</td><td>54</td><td>54</td><td>45</td><td>14</td><td>54</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>SpeⅠ</td>
| |
| - | <td>1</td><td>0</td><td>0</td><td>0</td><td>1</td><td>0</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>PstⅠ</td>
| |
| - | <td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>XbaⅠ</td>
| |
| - | <td>0</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0</td><td>0.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA(x10)</td>
| |
| - | <td>0.6</td><td>7</td><td>7</td><td>6</td><td>2</td><td>7</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Buffer2</td>
| |
| - | <td>6</td><td>-</td><td>-</td><td>-</td><td>2</td><td>-</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Buffer3</td>
| |
| - | <td>-</td><td>7</td><td>7</td><td>6</td><td>-</td><td>7<td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>1.9</td><td>1</td><td>1</td><td>2</td><td>0.5</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>60</td><td>70</td><td>70</td><td>60</td><td>20</td><td>70</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>----->37℃ 3hour
| |
| - | <br>----->Gel Extraction
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td><td>⑥</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA(digestion sample)</td>
| |
| - | <td>6</td><td>10</td><td>10</td><td>10</td><td>4</td><td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye</td>
| |
| - | <td>2</td><td>2</td><td>2</td><td>2</td><td>1</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>4</td><td>-</td><td>-</td><td>-</td><td>1</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>12</td><td>12</td><td>12</td><td>12</td><td>6</td><td>12</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>well</td>
| |
| - | <td>10</td><td>7</td><td>7</td><td>6</td><td>5</td><td>7</td>
| |
| - | </table>
| |
| - | <br>->DNA purification->①-④,⑥->7μl ⑤->6μl
| |
| - |
| |
| - | ----
| |
| - | '''8,Septmeber,2008'''
| |
| - | <br>SAP
| |
| - | <br>BBa_R0040(ptet)①
| |
| - | <br>BBa_R0010(pLac)⑤
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①</td><td>⑤</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA(digestion sample)</td>
| |
| - | <td>7</td><td>6</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>SAP/td>
| |
| - | <td>2</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>SAP Buffer</td>
| |
| - | <td>3</td><td>3</td><td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>30</td><td>30</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>30</td><td>30</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>->37℃ 1hour
| |
| - | <br>->65℃ 30min
| |
| - | <br>->DNA purification
| |
| - | <br>->21μl 6μl
| |
| - | <br>->Gel cheack
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>each</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA(digestion sample)</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye </td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>4</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>6</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>vector:BBa_R0040(ptet)① 75ng/μl
| |
| - | <br>insert:BBa_S03158(RBS+RhlR)② 33.6ng/μl
| |
| - | <br>insert:BBa_S03156(RBS+lasR)③ 33.6ng/μl
| |
| - | <br>insert:BBa_S03160(RBS+cinR)④ 38.4ng/μl
| |
| - | <br>vector:BBa_R0010(pLac)⑤ 100ng/μl
| |
| - | <br>insert:BBa_J52008(RBS+rLuc)⑥ 33.6ng/μl
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>①&②</td><td>①&③</td><td>①&④</td><td>①</td><td>⑤&⑥</td><td>⑤</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA(vector)</td>
| |
| - | <td>3</td><td>3</td><td>3</td><td>3</td><td>5</td><td>2.1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA(insert)</td>
| |
| - | <td>3</td><td>3</td><td>3</td><td>-</td><td>3.1</td><td>-</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Ligase</td>
| |
| - | <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Ligase Buffer(x5)</td>
| |
| - | <td>2</td><td>2</td><td>2</td><td>2</td><td>2</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>1</td><td>1</td><td>1</td><td>4</td><td>8.9</td><td>4</td>
| |
| - | </tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>10</td><td>10</td><td>10</td><td>10</td><td>20</td><td>10</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | ----
| |
| - | '''9,September,2008'''
| |
| - |
| |
| - | ----
| |
| - | '''10,September,2008'''
| |
| - | <br>TIME Responce(liquid)
| |
| - | <br>TIME Responce(solid)
| |
| - | <br>Colony PCR
| |
| - | <br>->Gel cheack
| |
| - | <br>MINI Prep
| |
| - |
| |
| - | ----
| |
| - | '''13,September,2008'''
| |
| - | <br>
| |
| - |
| |
| - |
| |
| - | ----
| |
| - | '''14,September,2008'''
| |
| - | <br>Digestion test
| |
| - | <br>BBa_Q04510
| |
| - | <br>BBa_I716463
| |
| - | <br>BBa_C0051
| |
| - | <br>BBa_C0070
| |
| - | <br>BBa_J06620
| |
| - | <br>BBa_J22141
| |
| - | <br>BBa_C0076
| |
| - | <br>BBa_I716542
| |
| - | <br>BBa_I716462
| |
| - | <br>BBa_C0078
| |
| - | <br>BBa_J01101
| |
| - | <br>BBa_C0071
| |
| - | <br>BBa_I712019
| |
| - | <br>BBa_S01416
| |
| - | <br>BBa_C0079
| |
| - | <br>BBa_C0077
| |
| - | <br>BBa_J23007
| |
| - | <br>BBa_I7100(x2)
| |
| - | <br>BBa_S01416
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>each</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA(digestion sample)</td>
| |
| - | <td>3</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Buffer2(x10)</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>6</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>EcoRⅠ</td>
| |
| - | <td>0.075</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>10</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>->37℃ 1hour
| |
| - | <br>Gel cheack
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>each</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA(digestion sample)</td>
| |
| - | <td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye</td>
| |
| - | <td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>12</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | ----
| |
| - | '''16,September,2008'''
| |
| - | <br>Digestion test
| |
| - | <br>BBa_R0040(ptet)+BBa_S03158(RBS+RhlR)(x2)
| |
| - | <br>BBa_R0040(ptet)+BBa_S03156(RBS+lasR)(x5)
| |
| - | <br>BBa_R0040(ptet)+BBa_S03160(RBS+cinR)(x2)
| |
| - | <br>BBa_S03158(RBS+RhlR)
| |
| - | <br>BBa_S03156(RBS+lasR)
| |
| - | <br>BBa_S03160(RBS+cinR)
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>each</td>
| |
| - | <tr>
| |
| - | <td>DNA</td>
| |
| - | <td>3</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>EcoRⅠ</td>
| |
| - | <td>0.083</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>PstⅠ</td>
| |
| - | <td>0.083</td>
| |
| - | </tr>
| |
| - | <td>BSA(x10)</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Buffer3(x10)</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>10</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>->37℃ 1hour
| |
| - | <br>Gel cheack
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>each</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA</td>
| |
| - | <td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye</td>
| |
| - | <td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>12</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>TRANSFORATION
| |
| - | <br>BBa_J37019(LuxI+RBS+LuxR)->no colony
| |
| - | <br>BBa_J37003(RBS+LuxR)
| |
| - | <br>BBa_J37020(LuxI+LuxR+pLux+GFP)
| |
| - | <br>BBa_J37032(pLux+GFP)
| |
| - | <br>PCR
| |
| - | <br>Venus YFP(ptet+LuxR+YFP)
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>YFP</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Venus Fwd</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Rev (VR)</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Thermo pol Buffer</td>
| |
| - | <td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dNTP</td>
| |
| - | <td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Vent</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>68</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>100</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | ----
| |
| - | '''17,September,2008'''
| |
| - | <br>TRANSFORMATION
| |
| - | <br>BBa_I13500(RBS+GFP)
| |
| - | <br>BBa_J37019(Luxpr+LuxR)-->no colony
| |
| - | <br>MINI Prep
| |
| - | <br>BBa_J37033(RBS+LuxR)
| |
| - | <br>BBa_J37020(Luxpr+LuxR+LuxR+GFP)
| |
| - | <br>BBa_J37032(pLux+GFP)
| |
| - | <br>Digestion
| |
| - | <br>BBa_I7100(ptetR+RBS)
| |
| - | <br>BBa_S03119(ptet+LuxR)
| |
| - | <br>Ligation product(ptet+LasR)
| |
| - | <br>Ligation product(ptet+RhlR)
| |
| - | <br>Ligation product(ptet+cinR)
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>each</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA template</td>
| |
| - | <td>25</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>EcoRⅠ</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>PstⅠ</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA(x10)</td>
| |
| - | <td>4</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>EcoRⅠ Buffer</td>
| |
| - | <td>4</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>40</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <br>-->37℃ 1hour
| |
| - | <br>->Next day
| |
| - | ----
| |
| - | '''18,September,2008'''
| |
| - | <br>SAP 1μl
| |
| - | <br>->37℃ 1hour
| |
| - | <br>->30min 65℃
| |
| - | <br>Zymo ->25μl
| |
| - | <br>Ligation
| |
| - | <br>vector:BBa_I7100
| |
| - | <br>insert:BBa_S03119(ptet+LuxR),Ligation product(ptet+LasR,ptet+RhlR,ptet+cinR)
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>Vector&insert</td><td>Vector</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA(vector)</td>
| |
| - | <td>3.5</td><td>3.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA(insert)</td>
| |
| - | <td>7.5</td><td>-</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Ligase</td>
| |
| - | <td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Ligase Buffer</td>
| |
| - | <td>4</td><td>4</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>4</td><td>4</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>20</td><td>20</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <br>TRANSFORMATION
| |
| - | <br>BBa_J54103(RBS+GFP)
| |
| - | <br>BBa_P2006
| |
| - |
| |
| - | <br>Gel Cheack
| |
| - | <br>Ligation product(Venus YFP)
| |
| - | <br>BBa_J37033
| |
| - | <br>BBa_J37020
| |
| - | <br>BBa_J37032
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>each</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA</td>
| |
| - | <td>each</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>4</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>6</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | ----
| |
| - | '''19,September,2008'''
| |
| - | <br>colony PCR
| |
| - | <br>ptet+LuxR(x16)
| |
| - | <br>ptet+LasR(x7)
| |
| - | <br>ptet+RhlR(x5)
| |
| - | <br>ptet+cinR(x5)
| |
| - |
| |
| - |
| |
| - | ----
| |
| - | '''22,September,2008'''
| |
| - | <br>Time delay
| |
| - |
| |
| - | ----
| |
| - | '''23,September,2008'''
| |
| - | <br>Digestion test
| |
| - | <br>BBa_E0240
| |
| - | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>E0240</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA</td>
| |
| - | <td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>EcoRⅠ</td>
| |
| - | <td>0.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>PstⅠ</td>
| |
| - | <td>0.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA(x10)</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>EcoRⅠ Buffer(x10)</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>10</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <br>Digestion
| |
| - | <br>Vector:BBa_E0240
| |
| - | <br>Insert:BBa_R0062
| |
| - | <br>Vector:BBa_I7100
| |
| - | <br>Insert:BBa_R0062
| |
| - | <br>Insert:BBa_J37033
| |
| | | | |
| | | | |
"
