Week1-Day 4 – PCR Round 2

Last evening we left our next two DNA samples in the PCR machine for amplification.

Glad to say that this morning all looks great. Time to check out if it really has worked-Time to create the DNA Barcode or known to teachers as DNA Profiling!

The agarose gel is once again made to receipe and the wells prepared. The gel is then loaded into the electrophoresis kit and buffer added.

Dye which is picked up by UV light is added to the DNA samples and the positive and negative controls.

The samples are then loaded into the pipettes (totally absorbed with precision, every drip is paramount) and the samples are carefully loaded into the wells of the gel. The samples sink to the bottom of the buffer covered wells as the dye contains glycerol which increases the density of the samples (another application of density! WHY are we learning this Miss? We’ll never use it!!! (Sound familiar?)

Switch on the electricity and DNA will run through the gel.  Today we used an orange dye for variation.

The gel is then placed in the uv chamber and hey presto our results looked great-A decent DNA barcode.

Clare and I spent some time researching and studying-Its so great to have actual set time for study-we all need it! 

The lab work certainly propels the desire to know as much as possible.

Back to the lab for the preparation of our PCR Master Mix-the PCR solution which contains all the elements needed for the process to occur.  It involves very detailed precise measurement-careful transfer of solutions and very intense attention to the micro-pipettes. One mistake can ruin a weeks work or a precious sample.

Samples readied and loaded into the PCR machine for our last run-this time we are using the samples which we purified on our first day! Very interested to see what the gel yields tomorrow!

Have to say that PCR should be taught in Secondary Schools-it is a fundamental lab skill-it brings together the skills and knowledge that are left segmented and isolated in the students minds.  It would also teach them how to order their practicals-analytical skills-attention to detail. Most importantly its fun and you get a real sense of achievement when those results are visualised!

 

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