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<center> <h1 id="MainTitle"><b> Experiments </b></h1> </center> | <center> <h1 id="MainTitle"><b> Experiments </b></h1> </center> | ||
+ | |||
+ | <section> | ||
+ | <div class="container"> | ||
+ | <div class="row"> | ||
+ | <div class="col-md-12 col-sm-10"> | ||
+ | |||
+ | The protocol we created was subject to change in order to produce optimal results. | ||
+ | <br><br> | ||
+ | 1. Innoculate viable colony into a liquid culture | ||
+ | <br> | ||
+ | - Materials: LB media, dilution, micropipette and tips | ||
+ | <br><br> | ||
+ | 2. Miniprep/Nanodrop | ||
+ | <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 | ||
+ | <br><br> | ||
+ | 3. Digest | ||
+ | <br> | ||
+ | -Materials: miniprepped DNA, dH₂O, 10X RE-Mix, standard restriction enzyme, micropipettes and tips | ||
+ | <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> | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </section> | ||
Revision as of 13:24, 13 October 2016
Experiments
The protocol we created was subject to change in order to produce optimal results.
1. Innoculate viable colony into a liquid culture
- Materials: LB media, dilution, micropipette and tips
2. Miniprep/Nanodrop
-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
3. Digest
-Materials: miniprepped DNA, dH₂O, 10X RE-Mix, standard restriction enzyme, micropipettes and tips
4. Gel
-Materials: agarose gel (make one if necessary), 1X TAE Buffer, power supply, chamber and electrodes, ladder, micropipette and tips, DNA
5. Ligation
-Materials: vector, parts 1 and 2, ligase buffer, ligase, Antarctic phosphatase, microcentrifuge tube, ice, micropipette and tips
6. Transformation and Plate
-Materials: ice, ligation mixture, competent cells, incubator, LB media, microcentrifuge tubes, micropipette and tips
7. Colony PCR (screening)
-Materials: dH₂O, buffer, VF₂, VR, Q5 polymerase, dNTP, DNA dilution, micropipette and tips, PCR tubes, thermocycler, ice
8. Gel
-Materials: agarose gel (make one if necessary), 1X TAE Buffer, power supply, chamber and electrodes, ladder, micropipette and tips, DNA
1. Innoculate viable colony into a liquid culture
- Materials: LB media, dilution, micropipette and tips
2. Miniprep/Nanodrop
-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
3. Digest
-Materials: miniprepped DNA, dH₂O, 10X RE-Mix, standard restriction enzyme, micropipettes and tips
4. Gel
-Materials: agarose gel (make one if necessary), 1X TAE Buffer, power supply, chamber and electrodes, ladder, micropipette and tips, DNA
5. Ligation
-Materials: vector, parts 1 and 2, ligase buffer, ligase, Antarctic phosphatase, microcentrifuge tube, ice, micropipette and tips
6. Transformation and Plate
-Materials: ice, ligation mixture, competent cells, incubator, LB media, microcentrifuge tubes, micropipette and tips
7. Colony PCR (screening)
-Materials: dH₂O, buffer, VF₂, VR, Q5 polymerase, dNTP, DNA dilution, micropipette and tips, PCR tubes, thermocycler, ice
8. Gel
-Materials: agarose gel (make one if necessary), 1X TAE Buffer, power supply, chamber and electrodes, ladder, micropipette and tips, DNA