IGEM:Hong Kong HKUST/Investigations/ Plasmid Cloning of BFP Generator Using Standard Assembly: Difference between revisions

IGEM:Hong Kong HKUST/Investigations/ Plasmid Cloning of BFP Generator Using Standard Assembly (view source)

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-===Restriction Enzyme Digestion===
+===Restriction Enzyme Digestion==
-[[Image:Digestion,_ligation.png]]
+J61002-BBa_J23110 was cut by the restriction enzyme SpeI-HF and PstI-HF and serves as the vector in the ligation, while pSB3K3-BBa_K592023 was cut by XbaI and PstI and isolated as an insert. 300ng of J61002-BBa_J23110 plasmid was used for digestion and 500ng of pSB3K3-BBa_K592023 plasmid was used for digestion. The volume of each enzyme used was 0.2μL and the buffer used was CutSmart. The temperature for digestion was 37°C and the digestion time was 1 hour 30 minutes.
-'''Figure 1. Standard assembly of BBa_J23110 and BBa_K592023.'''
-As illustrated in Figure 1, J61002-BBa_J23110 was cut by the restriction enzyme SpeI-HF and PstI-HF and serves as the vector in the ligation, while pSB3K3-BBa_K592023 was cut by XbaI and PstI and isolated as an insert. 300ng of J61002-BBa_J23110 plasmid was used for digestion and 500ng of pSB3K3-BBa_K592023 plasmid was used for digestion. The volume of each enzyme used was 0.2μL and the buffer used was CutSmart. The temperature for digestion was 37°C and the digestion time was 1 hour 30 minutes.
 ===Gel Electrophoresis, Purification, and Nanodrop Test===