training kit for students (how PCR works)
Purpose of the kit is to demonstrate students following:
- How PCR method functions?
- Which are the components of PCR?
- To show an example of the PCR application and working!
This kit is going to show how PCR can be used to detect human DNA in oral sample. This will be done with isolated DNA provided with kit. It is also possible that instructors can use other human isolated DNA source, if they have access to it.
The kit has different parts:
- Tube A: It contains forward and back primers. (Question for student: what are primers?)
- Tube B: It is master mix containing different dNTP, polymerase and buffer. (Question for students: what is dNTP and its role during the PCR; what is polymerase and what is its role)
- Tube D1: a tube containing positive material to run the assay. How can be DNA isolated from oral swabs?
- Tube D2: a tube without any human DNA to show that there will be no results, if there is no DNA inside!
- Tube E: a tube containing gene ruler 100 to 1000 bp (questions will be what is gene ruler and what is its role during the PCR test)
- Tube F: a tube containing loading dye (why does one need loading dye)
Equipment and material needed to conduct the test:
- PCR thermocycler
- Pipette Tips
Gel should be prepared before running the assay. It will teach how to prepare the gel.
At the end of test: one can see the specific band for human DNA and there will be no band in negative control D2, where there is no human DNA. Task for student is to make photo of the band on gel agarose or make a hand drawing to show how the band looks like. How to read the results of a gel agarose band?
More Questions for students:
- How can one isolate DNA from oral swabs?
- Why does PCR need isolated DNA?
Recommendation: There is a possibility that students can use their own DNA from buccal swabs to show how PCR works. In order to do it, Genekam EDU002 (DNA isolation kit for training) can be used.