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[[File:PCR_Side view_wiki.jpg]] | [[File:PCR_Side view_wiki.jpg]] | ||
With the 3 PCR's assembled and ready on Friday evening, Saturday morning came where we followed a DNA extraction protocol using the solutions (Primers.. Master mix.. Buffers.ec.t..) which came from a PRC Kit. | |||
So we Extracted DNA from our Breakfast: | |||
[[File:Breakfast-wiki.jpg]] | |||
The protocol involved breaking the histones, and using a centrifuge several times to extract the DNA. | |||
[[File:Extract_brkFast_wiki.jpg]] | |||
[[File:BioLab_DNA_wiki.jpg]] | |||
[[File:Centrifuge1_wiki.jpg]] | |||
[[File:DNA_ex_mont_1_wiki.jpg]] | |||
[[File:4_DNA_ex_2.jpg]] | |||
[[File:Extraction_rounds.jpg]] | |||
[[File:Final_DNA_Sample.jpg]] | |||
[[File:All_DNA_Samples.jpg]] | |||
The final samples are a cocktail of: | |||
DNA Sample to be tested | |||
Primers (forward and revers) 'Mark very specific lines of DNA code, (species specific) | |||
Deoxynucleotide triphosphates (dNTPs) - the nucleotides adenine (ATP), guanine (GTP), cytosine (CTP), and thymine (TTP) - the Characters of the code which will be used by the polymerase to replicate the DNA code that the Primers locate.) | |||
The Thermophilic polymerase with a buffer. (works well at 70 centigrade) | |||
[[File:Loaded_PCR.jpg]] | |||
[[File:PCR_cycles_running.jpg]] | |||
PCR Starts its first cycle! | |||
1. 95 °C Denaturation step. First, you heat the DNA to a high temperature (95 °C) so that the two strands of genomic DNA, and later PCR DNA, separate. | |||
2. Annealing Step (at ~ 50 - 60 °C). Second, you reduce the temperature so that DNA primers bind to either end of the template that you want to amplify. It is important that you have two primers, one to bind to each strand of DNA. | |||
3. Extension Step. Third, you raise the temperature to about 70 °C to activate a DNA polymerase and elongate the primer with respect to the template strand. | |||
This sequence above is repeated about 35 times. This results in 34,359,738,368 copies of the starting DNA sample. | |||