Team:Valencia UPV/Notebook

18May
  • May 18, 2016

L kanamycin + 5 μL Rifampin) 1:1000 and incubate overnight at 28^C.

19May
  • May 19, 2016

L in a new culture medium.

20May
  • May 20, 2016

Agroinfiltration in Nicotiana benthamiana of C58 with dsRED.

06Jun
  • June 06, 2016

  • href=" T"> Take from the glycerinates of Goldenbraid Collection:


Plasmid GB Code
1 GB0015
2 GB0017
1 GB0019
2 GB0021
pUPD2 GB0307


L antibiotic) 1:1000. Incubate at 37^C overnight.

  • href=" E"> Experiment with snails:

t been correctly infiltrated and it seems that the expression level is low.
Let both N. benthamiana leafs with snails overnight at room temperature in separated boxes. We will observe if snails eat the leafs and if appears fluorescence.

15Jun
  • June 15, 2016

t eaten leafs enough so we have not been able to see fluorescence.

30Jun
  • June 30, 2016

Orange DNA Genome Extraction protocol href

  • href=" T"> Take from the glycerinates of GoldenBraid Collection:


Plasmid GB Code
href="Prom">Promoter 35s:Cas9:nopaline synthase terminator (Tnos) GB0639
Luciferase (Luc) in pUPD2 GB0096
Tnos in pUPD2 GB0037


01Jul
  • July 01, 2016

. Plasmid Mini Kit I, Q(capless) Spin of:

  • href=" "> Promoter 35s:Cas9 : Tnos


t succeed. Repeat Luc and Tnos cultures.


Primers IG16JUN01 and IG16JUN02 have arrived.

Finish orange DNA Genome Extraction and check DNA concentration with NanoDrop. Concentration is very low so extraction will be done again.

02Jul
  • July 02, 2016

. Plasmid Mini Kit I, Q(capless) Spin of:

  • href=" "> Luc in pUPD2

    • href=" "> Tnos in pUPD2


    • href=" C"> Check orange DNA genome concentration with NanoDrop.


    L)
    Clemenules1 3153.8
    Clemenules 2 4527.9


    • href=" P"> Perform a PCR to bind linker with luciferase:

    L) Program
    LuciferasepUPD 1 Temperature Time
    Buffer HF 10 98^C 5 minutes
    dNTPs 2 98^C 35x 30 seconds
    IG16JUN01 2.5 70^C 35x 30 seconds
    IG16JUN02 2.5 72^C 35x 1 minute 30 seconds
    Taq phusion 0.5 72^C 10 minutes


04Jul
  • July 04, 2016


Run electrophoresis gel of Clemenules DNA (agarose 1%). 45 mL agarose gel at 1% 0.45 g of agarose with 45 mL of TAE 1X. Voltage used is 100 V.

Rice DNA Genome Extraction Protocol href

Run electrophoresis gel of Luciferase PCR product. 45 mL agarose gel at 1% 0.45 g of agarose with 45 mL of TAE 1X. Voltage used is 100 V.

Ligate Reaction --> Linker:Luciferase into a pUPD2

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