Lab 6
Today we ran the gel electrophoresis of yesterday's R.E.D. to confirm the successful ligation of the GAPC gene from Arabidopsis, which we amplified through nested PCR, with the pJet1.2 plasmid.
The results were used to determine which plasmid DNA purified samples were to be used in lab 7, when setting the sequencing reactions for the GAPC gene.
Lab 7
We added forward and reverse sequencing primers (pJET SEQ F and pJET SEQ R) to the plasmid DNA purified samples that had the GAPC gene insert 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 2-5 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.
(Note: Section 2 followed these protocols on Monday)
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