User:Samiye Yaman/Notebook/Lab 571/2011/11/01

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Entry title

Objective

Description

PCR

---Online webpage,( http://www.basic.northwestern.edu/biotools/oligocalc.html) was used to have a better understanding of PCR and find the forward and reverse primers necessary for the experiment.


Forward: 5' TAC GAC GAT GAC GAT AAG TGT CGA TGG GGA TCC GAA TTC 3'

Reverse: 3' ATG CTG CTA CTG CTA TTC ACA GCT ACC CCT AGG CTT AAG 5'

---A solution of 5 uL of 10X Pfu reaction buffer,1 uL of PfuTurbo DNA polymerase,1 uL of dNTP mix,1 uL of template DNA, 1.25 uL of primer 1 and 1.25 uL of primer 2 was prepared.

---Before placing the solution in the thermocycler at 95°C for 30 seconds, 25 uL of BioRad Chill out wax, dark pink, was added to the top of the solution in order to prevent the evaporation of the solution.

---The solution with the wax was kept in the thermocycler, which was at 4°C, for 24 hours.

Synthesizing gold nanoparticles.

  -Two test tubes were prepared.
  -Test tube A had 1 ml of HCl, 1 ml of BSA,  and 8 ml of water.(with this order)
  -Test tube B had 1 ml of BSA,7 ml of water and 2 ml of chloroauric acid.(with this order)
  -Both test tubes were placed in to the oven which was at 80 degree celcius and took it out of the oven 
    for 15 min every 30 minutes.

Data

PCR

Forward: 5' TAC GAC GAT GAC GAT AAG TGT CGA TGG GGA TCC GAA TTC 3'

Reverse: 3' ATG CTG CTA CTG CTA TTC ACA GCT ACC CCT AGG CTT AAG 5'


Synthesizing Goldnanoparticles

t=0 min

-Test tube A: clear color

-Test tube B: yellowish color


t:30 min

-Test tube A: clear solution

-Test tube B: yellow cloud at the top of the solution, bubbles in and out of the yellow cloud

t:60 min

-Test tube A: clear solution

-Test tube B: formation of black solid

t:120

-Test tube A:clear solution

-Test tube B: formation of black solid

t:150min

-Test tube A:clear solution

-Test tube B: formation of the black solid

t:180 min only test tube B was placed in to the oven. The black color of the solid became a dark red color.

Notes

This area is for any observations or conclusions that you would like to note.


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