Team:TCU Taiwan/Notebook

From 2014.igem.org

(Difference between revisions)
Line 223: Line 223:
<tr><td bgColor="#ffffff" colspan="3" height="10px"> </tr>
<tr><td bgColor="#ffffff" colspan="3" height="10px"> </tr>
<tr>
<tr>
-
   <td bgColor="#ffffff" colspan="3" width="70%"><fieldset>
+
   <td bgColor="#ffffff" colspan="3"><fieldset>
-
   <legend class="qnum" align="center"><font face="Trebuchet MS" size="6" color="#F24141">Gold</font><br>
+
  <legend class="qnum" align="center"><font face="Trebuchet MS" size="6" color="#F24141">Bacteria medium</font><br>
 +
  </legend>
 +
<div class="wrapper" style="background-color: white;">
 +
  <font size="3" face="Verdana" color="#333"><p>*LB(Luria-Bertani) broth: 1% Trytone, 1% NaCl, 0.5%  Yeast extract in the ddH2O<br>
 +
*LB  agar plate:LB broth add 1.5% agar.If you want to add selection marker example  for ampicillin or chloramphenicol. To culture <strong><em>E.coli</em></strong>, ampicillin is  added 100 mg/ml  for usage, chloramphenicol is added 30 mg/ml for usage,  kanamycin is added 70 mg/ml for usage</p>
 +
</font>
 +
</div>
 +
    </fieldset>
 +
</tr>
 +
<tr>
 +
  <td bgColor="#ffffff" colspan="3" height="10px">
 +
</tr>
 +
<tr>
 +
  <td bgColor="#ffffff" colspan="3" height="10px"><fieldset>
 +
  <legend class="qnum" align="center"><font face="Trebuchet MS" size="6" color="#F24141">Plasmid isolation</font><br></legend>
 +
  <div class="wrapper" style="background-color: white;">
 +
  <font size="3" face="Verdana" color="#333"><p>*Solution  I: 50 mM glucose、25 mM  Tris-HCl (pH 8.0)、10 mM EDTA<br>
 +
*Solution II:0.2N NaOH and 1% SDS(Sodium dodecyl sulfate&nbsp;)  mix<br>
 +
*Solution III: 5 M  potassium acetate (pH4.8), 11.5% glacial acetic acid<br>
 +
*RNase A(10 mg/ml)<br>
 +
*Phenol chloroform<br>
 +
*Chloroform<br>
 +
*95%  Ethanol<br>
 +
*Tris-EDTA(TE)buffer:10  mM Tris-Cl(pH 7.5) 1 mM EDTA<br>
 +
*pBluescript  SK(-)<br>
 +
*BBa_K1218011<br>
 +
*BBa_K914003<br>
 +
*BBa_I13521</p>
 +
</font>
 +
</div>
 +
 
 +
    </fieldset>
 +
</tr>
 +
<tr>
 +
  <td bgColor="#ffffff" colspan="3" height="10px">
 +
</tr>
 +
<tr>
 +
  <td bgColor="#ffffff" colspan="3" height="10px"><fieldset>
 +
  <legend class="qnum" align="center"><font face="Trebuchet MS" size="6" color="#F24141">Quick screening</font><br></legend>
 +
<div class="wrapper" style="background-color: white;">
 +
  <font size="3" face="Verdana" color="#333"><p>*Quick  screening buffer(Tris-Cl pH 12.6, 3% SDS)<br>
 +
*Phenol  chloroform<br>
 +
*Chloroform</p>
 +
</font>
 +
</div>
 +
    </fieldset>
 +
</tr>
 +
<tr>
 +
  <td bgColor="#ffffff" colspan="3" height="10px">
 +
</tr>
 +
<tr>
 +
  <td bgColor="#ffffff" colspan="3" height="10px"><fieldset>
 +
  <legend class="qnum" align="center"><font face="Trebuchet MS" size="6" color="#F24141">Enzyme digestion</font><br>
 +
</legend>
 +
<div class="wrapper" style="background-color: white;">
 +
  <font size="3" face="Verdana" color="#333"><p>*T4  ligase<br>
 +
*10X  ligation buffer<br>
 +
*<em>Eco</em>RI<br>
 +
*<em>Pst</em>I<br>
 +
*<em>Spe</em>I<br>
 +
*<em>Xba</em>I<br>
 +
*BSA  0.1%</p>
 +
</font>
 +
</div>
 +
    </fieldset>
 +
</tr>
 +
<tr>
 +
  <td bgColor="#ffffff" colspan="3" height="10px">
 +
</tr>
 +
<tr>
 +
  <td bgColor="#ffffff" colspan="3" height="10px"><fieldset>
 +
   <legend class="qnum" align="center"><font face="Trebuchet MS" size="6" color="#F24141">Bacteria strains and others</font><br>
   <font size="3" face="Verdana" color="#333"></font></legend>
   <font size="3" face="Verdana" color="#333"></font></legend>
<div class="wrapper" style="background-color: white;">
<div class="wrapper" style="background-color: white;">
-
   <p>&nbsp;</p>
+
   <p>*DH5a<br>
 +
    *JM101<br>
 +
    *0.8%  and 2% agarose gel<br>
 +
    *95%  Ethanol<br>
 +
    *6X  loading dye<br>
 +
    *1.5  ml centrifuge tube<br>
 +
    *250  ml flask<br>
 +
    *M13KO7(1*1011  pfu/ml)from <em>NEW ENGLAND BioLabs</em></p>
 +
</div>
 +
    </fieldset>
 +
</tr>
 +
<tr>
 +
  <td bgColor="#ffffff" colspan="3" height="10px">
 +
</tr>
 +
<tr>
 +
  <td bgColor="#ffffff" colspan="3" height="10px"><fieldset>
 +
  <legend class="qnum" align="center"><font face="Trebuchet MS" size="6" color="#F24141">Polymerase chain reaction</font><br></legend>
 +
<div class="wrapper" style="background-color: white;">
 +
  <font size="3" face="Verdana" color="#333"><p>*<em>Top</em> DNA polymerase<br>
 +
*dNTP(dATP,  dCTP, dGTP, dTTP) 250 mM<br>
 +
*Tris-HCl(pH9.0)  10 mM<br>
 +
*KCl  30 mM<br>
 +
*MgCl2  1.5 mM</p>
 +
</font>
 +
</div>
 +
    </fieldset>
 +
</tr>
 +
<tr>
 +
  <td bgColor="#ffffff" colspan="3" height="10px">
 +
</tr>
 +
<tr>
 +
  <td bgColor="#ffffff" colspan="3" height="10px"><fieldset>
 +
  <legend class="qnum" align="center"><font face="Trebuchet MS" size="6" color="#F24141">Instruments</font><br></legend>
 +
<div class="wrapper" style="background-color: white;">
 +
  <font size="3" face="Verdana" color="#333"><p>*VCM-620, laminal flow<br>
 +
*Biofuge pico, Heraeus<br>
 +
* OS1500-H, TSK Oribital Shaking Iucubator<br>
 +
* Biowave<br>
 +
* Vortex-GENE 2<br>
 +
* Autoclave AS-1060L<br>
 +
* GenAmp PCR System 2400, Applied  Biosystems<br>
 +
*BU-420,  YIHDER<br>
 +
*5804R,  eppendorf Centrifuge</p>
 +
</font>
</div>
</div>
     </fieldset>
     </fieldset>

Revision as of 18:08, 17 October 2014

 
Notebook
  
iGEM materials
Bacteria medium

*LB(Luria-Bertani) broth: 1% Trytone, 1% NaCl, 0.5% Yeast extract in the ddH2O
*LB agar plate:LB broth add 1.5% agar.If you want to add selection marker example for ampicillin or chloramphenicol. To culture E.coli, ampicillin is added 100 mg/ml for usage, chloramphenicol is added 30 mg/ml for usage, kanamycin is added 70 mg/ml for usage
Plasmid isolation

*Solution I: 50 mM glucose、25 mM Tris-HCl (pH 8.0)、10 mM EDTA
*Solution II:0.2N NaOH and 1% SDS(Sodium dodecyl sulfate ) mix
*Solution III: 5 M potassium acetate (pH4.8), 11.5% glacial acetic acid
*RNase A(10 mg/ml)
*Phenol chloroform
*Chloroform
*95% Ethanol
*Tris-EDTA(TE)buffer:10 mM Tris-Cl(pH 7.5) 1 mM EDTA
*pBluescript SK(-)
*BBa_K1218011
*BBa_K914003
*BBa_I13521
Quick screening

*Quick screening buffer(Tris-Cl pH 12.6, 3% SDS)
*Phenol chloroform
*Chloroform
Enzyme digestion

*T4 ligase
*10X ligation buffer
*EcoRI
*PstI
*SpeI
*XbaI
*BSA 0.1%
Bacteria strains and others

*DH5a
*JM101
*0.8% and 2% agarose gel
*95% Ethanol
*6X loading dye
*1.5 ml centrifuge tube
*250 ml flask
*M13KO7(1*1011 pfu/ml)from NEW ENGLAND BioLabs

Polymerase chain reaction

*Top DNA polymerase
*dNTP(dATP, dCTP, dGTP, dTTP) 250 mM
*Tris-HCl(pH9.0) 10 mM
*KCl 30 mM
*MgCl2 1.5 mM
Instruments

*VCM-620, laminal flow
*Biofuge pico, Heraeus
* OS1500-H, TSK Oribital Shaking Iucubator
* Biowave
* Vortex-GENE 2
* Autoclave AS-1060L
* GenAmp PCR System 2400, Applied Biosystems
*BU-420, YIHDER
*5804R, eppendorf Centrifuge
Life Records
   
 
 
 
Study Together
 
 
 
Conference in NCTU
 
 
Trip in Chihsingtan Beach
 
 
 
 
^


    
Team Members Project Parts Human Pratics Modeling Safety Notebook Attributions

Lost the way? Use it to help you if you're lost.

^

Retrieved from "http://2014.igem.org/Team:TCU_Taiwan/Notebook"