PCR!!

If only it was PBR right? (: (Professional Bull Riding) but no. This is almost as interesting, without the screaming fans, angry animals, and cowboys.


I did an online Bacterial ID Lab that was intended to identify the bacterial sample given.

First of all you have to prepare the DNA sample that is to be used. This involves placing the bacterial colony in a microcentrifuge tube and inserting the digestive buffer. The tube has to sit for several hours until the sample is ready. Following that you have to heat-inactivate the digestive enzymes which will denature and inactivate them. Add counterbalance and spin down cellular debris for removal from the sample. Then the wanted DNA is supernatant. Transfer the supernatant to the PCR tube and the sample prep is complete!!



Next step is to use PCR to make copies of DNA sequences. To do this you have to add a PCR master mix solution to all of the tubes. To set up a positive control reaction, you have to add control dna to one tube. For negative reactions you must add de-ionized water to one tube. Then insert both of them into the PCR machine. The short strands are desired DNA strands from PCR thermal cycling process.



NEXT: Purify the PCR product. To do this you have to set up the microconcentrator column and add a buffer solution to the column, then add the PCR product to the column. Put the positive and negative controls on ice until needed again. Add counterbalance to the tube and put it in the centrifuge. After a while DNA is trapped into the column and you need to loosen it into a new tube. Invert the column into the new tube (turn it upside down) and add a buffer. Now put centrifuge assembly to collect DNA into a collection and wait patiently.



PCR sequencing prep is next. First you have to dillute the PCR product to increase the volume. Add distilled water to purify the PCR product. The green and blue strip tubes contain PCR reaction mixtures, so now you can add purifed PCR product to them. They are now ready for PCR amplification so put it in the PCR machine. The Sequencing prep is complete!



Second to last step is DNA Sequencing. You must load the auto sequencer and insert the tubes. In my case the first tube read tacggtagcgtaatgcc. And that completes the DNA sequencing event.



The last step is to analyze the sequence. Copy the output from the DNA sequencer and past it on the ncbi website. The ncbi website searches through it's massive database and eventually comes up with results. The bacteria was identified as bartonella henselae.