Team:Vanderbilt/Notebook

• Extracted genomic DNA from 100 mg of Arabidopsis thaliana leaves using a Viogene DNA extraction kit. Samples were labeled ZIN for zingiberene.
• Two samples were prepared and nanodropped. The concentration of the first was 2.6 ng/ul, and the second was 3.5 ng/ul of DNA, indicating a minimal yield of DNA.

• Extracted genomic DNA from 100 mg of Picea abies needles using the same extraction kit and protocol. Samples were labeled CAR for carene.
• Two samples were prepared and nanodropped. The concentration of the first was 0.9 ng/ul, and the second was 3.5 ng/ul of DNA, indicating a minimal yield of DNA.

• Ran a 0.6 % argarose gel on the DNA extracted from ZIN and CAR, as well as the column flow-through from the kit.
• ZIN1, ZIN2, CAR1, and CAR2 all show a faint but distinct DNA band above the highest rung on the DNA ladder (>10kb), showing the presence of DNA. No bands were seen on the kit flow-through. Indicates successful genomic extraction.
• Preformed a second genomic extraction on Picea abies to improve yield. Nanodrop shows CAR3 to be at 6.2 ng/ul and CAR4 to be 11.5 ng/ul.
• Extracted genomic DNA from Gossypium hirsutum . Samples nanodroppped: CAD1 1.8 ng/ul and CAD2 7.8 ng/ul.

• Ran a gel on CAR3, CAR4, CAD1, and CAD2. Brighter genomic DNA bands were seen on the cadinene camples than before, but cadinene samples showed significant smearing near the top of the gel.

• Extracted genomic DNA from Salvia officinalis . Samples nanodropped: SAB1 7.5 ng/ul, and SAB 11.4 ng/ul.
• Extracted genomic DNA from Mentha citrata . Samples nanodropped: LNR1 3.4 ng/ul, LNR2 6.2 ng/ul, LNR3 7.3 ng/ul
• E. coli containing p404GALS and pDZ207 from Addgene were grown on LB plates with ampicilin. These were miniprepped using a Viogen kit. Samples nanodropped: p404GALS (A) 130.2 ng/ul, p404GALS (B) 112.3 ng/ul, pDZ207 (A) 145.3 ng/ul, pDZ207 (B) 84.1 ng/ul.

• Extracted genomic DNA again from Arabidopsis thaliana . Sample nanodropped: LNS1 53.4 ng/ul, LNS2 585.2 ng/ul
• Ran gel on LNR1, LNR2, LNR3, LNS1, LNS2, SAB1, and SAB2. All show high weight DNA bands, with Linalool (S) samples having the brightest.

• Extracted genomic DNA from Zingiber zermbet . Sample nanodropped: HUM1 31.9 ng/ul and HUM2 18.6 ng/ul
• Extracted genomic DNA from Ocimum basilicum . Sample nanodropped: GER1 2.3 ng/ul

• Extracted genomic DNA from Santalum album seeds since the sapling was still not fully grown. Sample nanodropped: SAN1 53.8 ng/ul and SAN2 28.4 ng/ul

• Extracted genomic DNA from Perilla frutescens . Sample nanodropped: MYR1 632.8 ng/ul and MYR2 958.6 ng/ul
• Ran gel on MYR1, MYR2, SAN1, and SAN2. Myrcene samples show bright band at high weight as well as smearing toward bottom. Santelene samples have visible but much fainter genomic DNA band.

• Preformed PCR using zingiberene synthase primers on LNS2 genomic DNA and using linalool (S) synthase primers on the sample template.
• Ran gel on PCR product. Resulted in no visible bands formed.

• Took 1.8 ul of HUM1 genomic DNA and preformed a PCR with humelene synthase primers.
• Ran gel on PCR product. Resulted in three total bands, one faint around 1.2 kb, and two bands very close in size just under 3.0 kb.
• Both ~3 kb bands were gel extracted, combining across all four lanes. Samples nanodropped: HUM-top 8.5 ng/ul, HUM-bottom 10.6 ng/ul.

• Preformed PCR using linalool (R) synthase primers on LNR3 genomic DNA.
• Ran gel on PCR product. Resulted in no visible bands formed.

• Preformed PCR using myrcene synthase primers on MYR2 genomic DNA.
• Ran gel on PCR product. Smeared bands on gel but no distinct bands.
• Preformed overlap-extension PCR on the gel extracted humelene samples (both HUM-top and HUM-bottom) to add epitope tag sequence
• Ran gel on OE-PCR product. Only extremely faint bands were visible.

Summer 2014