Team:UNIPV-Pavia/Calendar/July/settimana2

July, 4th

Digestions of previously purified plasmids were performed for ligations:

Plasmid	Kind	Purified Dna (μl)	H2O (μl)	Enzyme 1 (μl)	Enzyme 2 (μl)	Buffer H (μl)	Final Volume(μl)
<partinfo>BBa_C0060</partinfo>	Insert	16.5	4	1 EcoRI	1 SpeI	2.5	25
<partinfo>BBa_C0061</partinfo>	Insert	13.2	7.3	1 XbaI	1 PstI	2.5	25
<partinfo>BBa_K081022</partinfo>	Insert	15.7	4.8	1 EcoRI	1 PstI	2.5	25
<partinfo>BBa_B0030</partinfo>	Vector	13.2	7.3	1 SpeI	1 PstI	2.5	25
<partinfo>BBa_B0031</partinfo>	Vector	12.4	8.1	1 SpeI	1 PstI	2.5	25
<partinfo>BBa_B0032</partinfo>	Vector	9.5	11	1 SpeI	1 PstI	2.5	25
<partinfo>BBa_B0015</partinfo>	Vector	7.9	12.6	1 EcoRI	1 XbaI	2.5	25
<partinfo>pSB4C5</partinfo>	Vector	3	17.5	1 EcoRI	1 PstI	2.5	25
<partinfo>BBa_I13501</partinfo>	Insert	7.2	13.3	1 XbaI	1 PstI	2.5	25

Reaction samples were incubated at 37°C for three hours while a small-size and a medium-size agarose gel were prepared according to protocols.

In the afternoon gel electrophoresis was performed.

[Image:UNIPV_04_07_SSgel.png]

As shown, in figure all clones were positive (apart from <partinfo>BBa_I13501</partinfo> run where we inexplicably didn't see any band), so we cut and purified the bands of interest.

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