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1.Prepare RNA mix in PCR tube 1.1 µg RNA 2.1 µl Random primer/poly dT mix 3.1 µl 10 mM dNTP 4.Water to 12 µl 2.Leave 65˚ C, then chill to 4˚ C 3.Add.

Published byMeghan Lyons Modified over 8 years ago

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Presentation on theme: "1.Prepare RNA mix in PCR tube 1.1 µg RNA 2.1 µl Random primer/poly dT mix 3.1 µl 10 mM dNTP 4.Water to 12 µl 2.Leave 65˚ C, then chill to 4˚ C 3.Add."— Presentation transcript:

1 1.Prepare RNA mix in PCR tube 1.1 µg RNA 2.1 µl Random primer/poly dT mix 3.1 µl 10 mM dNTP 4.Water to 12 µl 2.Leave 5’ @ 65˚ C, then chill to 4˚ C 3.Add 1.4 µl 5x first strand buffer 2.2 µl 100 mM DTT 3.1 µl RNAse inhibitor 4.Leave > 2’ @ RT 5.Add 1 µl Superscript III 6.Leave 10’ @ 25 ˚ C, then 50’ @ 42 ˚ C 7.Inactivate by leaving 15’ @ 70˚ C 8.Use 1 µl for PCR with gene-specific primers

2 1.Set up master mix for each primer combo on ice! 1.1.1 µl 100x F primer (1 pMol/µl = 1µM final []) 2.1.1 µl 100x R primer 3.11 µl 10x PCR buffer 4.2.2 µl 10 mM dNTP (200 µM final []) 5.88.4 µl water 6.0.7 µl Taq polymerase 2.Primers are 1.corn SOD or PEPC 2.Rice OEE or PHYA 3.Arabidopsis Actin and Aromatic Acid Decarboxylase 3.Add 19 µl to 1 µl correct cDNAs, 19 µl to 1 µl correct genomic DNA, 19 µl to 1 µl no template cDNA and 19 µl to 1 µl water 4.Run 30 cycles of 15” @ 94, 50-1”/cycle @ 50, 15” @ 72

3 Possible Projects 1.Studying interactions between plants and Geobacter (or other electrogenic bacteria)

4 Possible Projects 1.Studying interactions between plants and Geobacter (or other electrogenic bacteria) 2.Studying interactions between cyanobacteria and Geobacter (or other electrogenic bacteria)

5 Possible Projects 1.Studying interactions between plants and Geobacter (or other electrogenic bacteria) 2.Studying interactions between cyanobacteria and Geobacter (or other electrogenic bacteria) 3.Studying potential of Rhodopseudomonas (a.k.a. Rhodobacter) for biophotovoltaics

6 Possible Projects 1.Studying interactions between plants and Geobacter (or other electrogenic bacteria) 2.Studying interactions between cyanobacteria and Geobacter (or other electrogenic bacteria) 3.Studying potential of Rhodopseudomonas (a.k.a. Rhodobacter) for biophotovoltaics Switch to photosynthesis when go anaerobic!

7 Possible Projects 1.Studying interactions between plants and Geobacter (or other electrogenic bacteria) 2.Studying interactions between cyanobacteria and Geobacter (or other electrogenic bacteria) 3.Studying potential of Rhodopseudomonas (a.k.a. Rhodobacter) for biophotovoltaics Switch to photosynthesis when go anaerobic! Selection for cells that can transfer e- to electrodes!

8 Possible Projects 1.Studying interactions between plants and Geobacter (or other electrogenic bacteria) 2.Studying interactions between cyanobacteria and Geobacter (or other electrogenic bacteria) 3.Studying potential of Rhodopseudomonas (a.k.a. Rhodobacter) for biophotovoltaics 4.Studying Cyanobacteria for biophotovoltaics

9 Possible Projects 4. Studying Cyanobacteria for biophotovoltaics Finding new ones

10 Possible Projects 4. Studying Cyanobacteria for biophotovoltaics Finding new ones Tweaking knowns http://www.invitrogen.co m/site/us/en/home/Produ cts-and- Services/Applications/Pr otein-Expression-and- Analysis/Protein- Expression/algae- engineering-kits.html

11 Possible Projects 4. Studying Cyanobacteria for biophotovoltaics Finding new ones Tweaking knowns Tweaking nanowires

12 Possible Projects 5. Studying green algae for biophotovoltaics Finding new ones Tweaking knowns (Chlorella, Dunaliella)

13 Possible Projects 6. Studying plants for biophotovoltaics Roots under hypoxia

14 Possible Projects 6. Studying plants for biophotovoltaics Roots under hypoxia Aquatics under CO 2 deprivation

15 Possible Projects 6. Studying plants for biophotovoltaics Roots under hypoxia Aquatics under CO 2 deprivation = every day!

16 Possible Projects 6. Studying plants for biophotovoltaics Roots under hypoxia Aquatics under CO 2 deprivation = every day! Some aquatics do CAM because CO 2 is low in water in the day

17 Possible Projects 6. Studying plants for biophotovoltaics Roots under hypoxia via mediator Aquatics under CO 2 deprivation Some aquatics do CAM because CO 2 is low in water in the day Perhaps others will transfer electrons to anodes during the day to dissipate excess energy

18 Possible Projects 7. Engineering the production of other novel products

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