User:Samiye Yaman/Notebook/Lab 571/2011/09/21

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Entry title

Objective

Description

Procedures from the day before was continued

1)A culture media of E.Coli was prepared for the protein expression.

-The solutions with Ecoli cultures from each flask transferred into 4 different bottles for centrifuging.

-After making sure that each bottle had similar weight, they were placed into centrifuge at 4500rpm for 50 minutes at 4°C.

-After the centrifuge, the solution was discarded and the solid at the bottom of each bottle was obtained by using 200 ml of column buffer which was made up of 20mM trish HCL, 200Mm NaCl and 1mM EDTA and placed into test tubes.


2)GFP containing a cysteine was prepared by using PCR.

-The solution with the wax was was removed from thermocycler.

-By using pipette, the wax was removed from the solution.

-By suing 0.25g of agarose and 25 mL of TAE buffer, 0.01g/mL agarose gel was prepared. There were particles floating in the solution

-The agarose gel was placed into the microwave for 40 seconds in order to have a homogeneous solution.

-The clear homogeneous solution was placed into a mold for 15 min to cool down and solidify.

- More TAE buffer was poured to the top of the agarose gel when it became solid.

-1 ul of blue dye and glycerol mixture and 5uL of PCR were mixed well before loading them into the gel.

-80 V was passed through the gel for 40 min.

Data

  • Add data and results here...

Image:DNA_gel_092111_003.jpg

Notes

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