UCLAiGEM:Notebook/MichaelCheng/2015/01/28

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Digestion of pSB1a3 and Tamura insert

pSB1a3 and Tamura digested with EcoRI and PstI.


Digestion Recipe: Tamura
Material Amount
EcoRI 1 uL
PstI 1 uL
10x NEB 2.1 Buffer 5 uL
"Tamura #2" 1 ug DNA
Water fill to 50 uL


Digestion Recipe: pSB1a3
Material Amount
EcoRI 1 uL
PstI 1 uL
10x NEB 2.1 Buffer 5 uL
"Tamura Miniprep 1" 1 ug DNA
Water fill to 50 uL

Notice: This is confusing as all hell: Tamura Miniprep 1 is what we are digesting to obtain the linearized and digested pSB1a3. Turns out, we don't have any actual linear pSB1a3, so we have to obtain it from a completed Tamura miniprep that we performed ages ago.


Running the digested Tamura and pSB1a3 on a gel

Gel was 1% w/v, 40 mL. Was run for 30 minutes at 100 V.


The bands containing Tamura and pSB1a3 were then excised and stored. Tamura: Serial# D00008 pSB1a3: Serial# D00007





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