Module 2, Lab 07 - Sequencing reactions of GAPC gene

After the cloning process of the GAPC gene from Arabidopsis, and extracting the plasmid DNA (to isolate the pJet1.2 plasmid) we did a RED to confirm the success of the ligation.

Using the samples that had the GAPC gene insert we mixed purified plasmid DNA with forward and reverse sequencing primers (pJET SEQ F and pJET SEQ R), and put them in a 96-well plate. The plate will be shipped to the DOE Joint Genome Institute (JGI) to be sequenced as part of their Sequencing Training Program (STR). The results should be in in two weeks, ready to be used in the bioinformatics labs

We will discuss the DNA sequencing technique most commonly used: Dye-terminator sequencing, a modification of Sanger's chain termination sequencing protocol, which allowed the automation of the DNA sequencing process.

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