IGEM:UNAM-Genomics Mexico/2009/Notebook/H2 Pae/2011/06/28

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Results of the digestion's test

The following gel(120V x 35 min) shows the results of the pBBRMCS-5's digestion by apaI, which was made yesterday:

Lanes:
1. Ladder 10Kb.
2. Digestion.
3. pBBMRCS-5 isolation.

Pae 28 jun 2011.png

Here we clearly see that the isoforms in lane 3 doesn`t exit in the lane 2, besides the band corresponds to the correct size of the plasmid (arround 4.6 Kb). With this we can proceed to double digestion (apaI, sacI).

Double digestion

Paulina did the pBBRMCS-5's digestion only with sacI to see if it's working right. The reagents ratio is :

H2O 2 microL
Buffer 4 1 microL
BSA 1 micrL
sacI 1 microL
DNA 5 microL
Total 10 microL

I made the double digestion with apaI and sacI, which will be ligated to a fragment containing prefix-RFP-suffix.

The ratio:

Buffer 4 2 microL
BSA 2 microL
sacI 1.5 microL
apaI 1.5 micrL
DNA 13 microL
Total 20 microL

Tomorrow we'll see the results in an electrophoresis gel.