IGEM:UNAM/2009/Notebook/IGEM project/2011/07/26

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Gel of plasmid and digestion

  • Today I did the gels of the last extraction.

4. Gel of Plasmid (20 lines)

Line 1 -> Green ladder 2μl.
Line 2 -> 3 μl of plasmid J23112A + 1 μl de dye.
Line 3 -> 3 μl of plasmid J23113A + 1 μl de dye.
Line 4 -> 3 μl of plasmid J23114A + 1 μl de dye.
Line 5 -> 3 μl of plasmid J23115A + 1 μl de dye.
Line 6 -> 3 μl of plasmid J23116A + 1 μl de dye.
Line 7 -> 3 μl of plasmid J23117A + 1 μl de dye.
Line 8 -> 3 μl of plasmid J23118A + 1 μl de dye.
Line 9 -> 3 μl of plasmid J23119A + 1 μl de dye.
Line 10 -> 3 μl of plasmid J23112B + 1 μl de dye.
Line 11 -> 3 μl of plasmid J23113B + 1 μl de dye.
Line 12 -> 3 μl of plasmid J23114B + 1 μl de dye.
Line 13 -> 3 μl of plasmid J23115B + 1 μl de dye.
Line 14 -> 3 μl of plasmid J23116B + 1 μl de dye.
Line 15 -> 3 μl of plasmid J23117B + 1 μl de dye.
Line 16 -> 3 μl of plasmid J23118B + 1 μl de dye.
Line 17 -> 3 μl of plasmid J23119B + 1 μl de dye.
Line 18 -> 3 μl of plasmid J23112C + 1 μl de dye.
Line 19 -> 3 μl of plasmid J23113C + 1 μl de dye.
Line 20 -> 3 μl of plasmid J23114C+ 1 μl de dye.

5. Gel of Plasmid (15 lines)

Line 1 -> Green ladder 2μl.
Line 2 -> 3 μl of plasmid J23115C + 1 μl de dye.
Line 3 -> 3 μl of plasmid J23117C + 1 μl de dye.
Line 4 -> 3 μl of plasmid J23118C + 1 μl de dye.
Line 5 -> 3 μl of plasmid J23119C + 1 μl de dye.
Line 6 -> 3 μl of plasmid J23112D + 1 μl de dye.
Line 7 -> 3 μl of plasmid J23113D + 1 μl de dye.
Line 8 -> 3 μl of plasmid J23114D + 1 μl de dye.
Line 9 -> 3 μl of plasmid J23115D + 1 μl de dye.
Line 10 -> 3 μl of plasmid J23117D + 1 μl de dye.
Line 11 -> 3 μl of plasmid J23119D + 1 μl de dye.
Line 12 -> 3 μl of plasmid J23115D + 1 μl de dye.
Line 13 -> Empty.
Line 14 -> Empty.
Line 15 -> Empty.

Besides I selected the best extraction to do the digestion:

J23100A, J23101A, J2312A, J23103A, J23104A, J23105A, J23106A, J23107A, J23108A, J23109A, J23110A, J23100B, J234101B, J23102B,
J23104B, J23105B, J2306B, J23108B, J2311!B, J23101C, J23102C, J23194C, J23198C, J23109C, J23109B, J23117A.
  • Digestion
Reagent Quantity
H2O miliq 6 μl
Buffer 10X of EcorRI 1 μl
plasmid 2 μl
EcoRI enzyme 1 μl

Total 10 μL. Wait 2:30 hours to 37 °C and make a gel to see if the digetion was made in the plasmid.

6. Gel of digestion (20 lines)

Line 1 -> Green ladder 2μl.
Line 2 -> 3 μl of digestion J23100A + 1 μl de dye.
Line 3 -> 3 μl of digestion J23101A + 1 μl de dye.
Line 4 -> 3 μl of digestion J23102A + 1 μl de dye.
Line 5 -> 3 μl of digestion J23103A + 1 μl de dye.
Line 6 -> 3 μl of digestion J23104A + 1 μl de dye.
Line 7 -> 3 μl of digestion J23105A + 1 μl de dye.
Line 8 -> 3 μl of digestion J23107A + 1 μl de dye.
Line 9 -> 3 μl of digestion J23106A + 1 μl de dye.
Line 10 -> 3 μl of digestion J23108A + 1 μl de dye.
Line 11 -> 3 μl of digestion J23109A + 1 μl de dye.
Line 12 -> 3 μl of digestion J23110A + 1 μl de dye.
Line 13 -> 3 μl of digestion J23100B + 1 μl de dye.
Line 14 -> 3 μl of digestion J23101B + 1 μl de dye.
Line 15 -> 3 μl of digestion J23102B + 1 μl de dye.
Line 16 -> 3 μl of digestion J23103B + 1 μl de dye.
Line 17 -> 3 μl of digestion J23104B + 1 μl de dye.
Line 18 -> 3 μl of digestion J23105B + 1 μl de dye.
Line 19 -> 3 μl of digestion J23106B + 1 μl de dye.
Line 20 -> 3 μl of digestion J23101C+ 1 μl de dye.

7. Gel of digestion (15 lines)

Line 1 -> Green ladder 2μl.

Line 2 -> 3 μl of digestion J23102C + 1 μl de dye.
Line 3 -> 3 μl of digestion J23109B + 1 μl de dye.
Line 4 -> 3 μl of digestion J23102A + 1 μl de dye.
Line 5 -> 3 μl of digestion J23104C + 1 μl de dye.
Line 6 -> 3 μl of digestion J23108B + 1 μl de dye.
Line 7 -> Empty.
Line 8 -> 3 μl of digestion J23111B + 1 μl de dye.
Line 9 -> 3 μl of digestion J23117A + 1 μl de dye.
Line 10 -> 3 μl of digestion J23109C + 1 μl de dye.
Line 11 -> 3 μl of digestion J23108C + 1 μl de dye.
Line 12 -> Empty.
Line 13 -> Empty.
Line 14 -> Empty.
Line 15 -> Empty.

At last in the night we grow Escherichia coli DH5-α to extract plasmid.



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