Difference between revisions of "Team:LambertGA/Experiments"

Line 366: Line 366:
 
<div class="col-md-12 col-sm-10" style="width:75%;margin:auto;">
 
<div class="col-md-12 col-sm-10" style="width:75%;margin:auto;">
  
−
The protocol we created was subject to change in order to produce optimal results.
+
<h3 style="text-decoration: none; color: #D49AE6;"> Workflow </h3>
 
<br><br>
 
<br><br>
−
1. Innoculate viable colony into a liquid culture
+
1. Miniprep/Nanodrop <br>
−
<br>
+
2. Digest <br>
−
- Materials: LB media, dilution, micropipette and tips
+
3. Gel <br>
−
<br><br>
+
4. Ligation <br>
−
2. Miniprep/Nanodrop
+
5. Transformation, Plate <br>
−
<br>
+
6. Colony PCR (Screening) <br>
−
-Materials: grown culture, microcentrifuge, 2 1.5mL microcentrifuge tubes, mini column and collection tube, Solution I, Solution II, Solution III, HBC Wash Buffer, DNA Wash Buffer, Elution Buffer, micropipette and tips
+
7. Gel <br>
−
<br><br>
+
8. Inoculate correct colony to a liquid culture. <br>
−
3. Digest
+
 
−
<br>
+
 
−
-Materials: miniprepped DNA, dH₂O, 10X RE-Mix, standard restriction enzyme, micropipettes and tips
+
<h3 style="text-decoration: none; color: #D49AE6;"> Materials: </h3>
−
<br><br>
+
−
4. Gel
+
−
<br>
+
−
-Materials: agarose gel (make one if necessary), 1X TAE Buffer, power supply, chamber and electrodes, ladder, micropipette and tips, DNA
+
−
<br><br>
+
−
5. Ligation
+
−
<br><br>
+
−
-Materials: vector, parts 1 and 2, ligase buffer, ligase, Antarctic phosphatase, microcentrifuge tube, ice, micropipette and tips
+
−
<br><br>
+
−
6. Transformation and Plate
+
−
<br><br>
+
−
-Materials: ice, ligation mixture, competent cells, incubator, LB media, microcentrifuge tubes, micropipette and tips
+
−
<br><br>
+
−
7. Colony PCR (screening)
+
−
<br><br>
+
−
-Materials: dH₂O, buffer, VF₂, VR, Q5 polymerase, dNTP, DNA dilution, micropipette and tips, PCR tubes, thermocycler, ice
+
−
<br><br>
+
−
8. Gel
+
−
<br><br>
+
−
-Materials: agarose gel (make one if necessary), 1X TAE Buffer, power supply, chamber and electrodes, ladder, micropipette and tips, DNA
+
 
<br><br>
 
<br><br>
 +
Miniprep: grown culture, microcentrifuge, 2 1.5mL microcentrifuge tubes, mini column and collection tube, Solution I, Solution II, Solution III, HBC Wash Buffer, DNA Wash Buffer, Elution Buffer, micropipette and tips<br>
 +
Nanodrop: nanodrop machine, miniprepped DNA, Kimtech wipes, micropipette and tips<br>
 +
Digest: miniprepped DNA, dH₂O, 10X RE-Mix, standard restriction enzyme, micropipettes and tips<br>
 +
Gel: agarose gel (make one if necessary), 1X TAE Buffer, power supply, chamber and electrodes, ladder, micropipette and tips, DNA<br>
 +
Ligation: vector, parts 1 and 2, ligase buffer, ligase, Antarctic phosphatase, microcentrifuge tube, ice, micropipette and tips <br>
 +
Transformation: ice, ligation mixture, competent cells, incubator, LB media, microcentrifuge tubes, micropipette and tips
 +
Plate: agar plate, micropipette and tips, beads<br>
 +
Colony PCR: dH₂O, buffer, VF₂, VR, Q5 polymerase, dNTP, DNA dilution, micropipette and tips, PCR tubes, thermocycler, ice<br>
 +
Gel: agarose gel (make one if necessary), 1X TAE Buffer, power supply, chamber and electrodes, ladder, micropipette and tips, DNA<br>
 +
Inoculate: LB media, dilution, micropipette and tips<br>
 +
 
</p>
 
</p>
 
</div>
 
</div>

Revision as of 23:38, 18 October 2016


Experiments


Workflow



1. Miniprep/Nanodrop
2. Digest
3. Gel
4. Ligation
5. Transformation, Plate
6. Colony PCR (Screening)
7. Gel
8. Inoculate correct colony to a liquid culture.

Materials:



Miniprep: grown culture, microcentrifuge, 2 1.5mL microcentrifuge tubes, mini column and collection tube, Solution I, Solution II, Solution III, HBC Wash Buffer, DNA Wash Buffer, Elution Buffer, micropipette and tips
Nanodrop: nanodrop machine, miniprepped DNA, Kimtech wipes, micropipette and tips
Digest: miniprepped DNA, dH₂O, 10X RE-Mix, standard restriction enzyme, micropipettes and tips
Gel: agarose gel (make one if necessary), 1X TAE Buffer, power supply, chamber and electrodes, ladder, micropipette and tips, DNA
Ligation: vector, parts 1 and 2, ligase buffer, ligase, Antarctic phosphatase, microcentrifuge tube, ice, micropipette and tips
Transformation: ice, ligation mixture, competent cells, incubator, LB media, microcentrifuge tubes, micropipette and tips Plate: agar plate, micropipette and tips, beads
Colony PCR: dH₂O, buffer, VF₂, VR, Q5 polymerase, dNTP, DNA dilution, micropipette and tips, PCR tubes, thermocycler, ice
Gel: agarose gel (make one if necessary), 1X TAE Buffer, power supply, chamber and electrodes, ladder, micropipette and tips, DNA
Inoculate: LB media, dilution, micropipette and tips