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Revision as of 20:28, 13 September 2013 (view source) Queziatoe (Talk | contribs) (→Gel Purification of the pGEM-T pasmid double digestions ran in the 11/09/13) ← Older edit		Revision as of 20:28, 13 September 2013 (view source) Queziatoe (Talk | contribs) (→Gel Purification of the pGEM-T pasmid double digestions ran in the 11/09/13) Newer edit →
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	|TOBG||47.7||9||1.77||0.70		|TOBG||47.7||9||1.77||0.70
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	|pSB1C3 (RFP)||46.2||7.3||1.81||0.16		|pSB1C3 (RFP)||46.2||7.3||1.81||0.16
	|}		|}

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Revision as of 20:28, 13 September 2013

Contents 1 Isolate plasmid from overnight culture 2 Glycerol stocks 3 Gel Purification of the pGEM-T pasmid double digestions ran in the 11/09/13 4 Digestion of the samples purified with the enzyme PstI-HF

Isolate plasmid from overnight culture

• Using Omega mini prep kit protocol
• Samples:
  • TODF 1,2,3,
  • TODX 1, 2, 3
  • TOBG 1, 2, 3,
  • RFP 1, 2, 3

sample	Volume (ul)	Concentration (ng/ul)
TODF1	43	120
TODF2	43	98.8
TODX1	42	399.3
TODX2	44	442.2
TODX3	43	417.5
TOBG1	43	262.4
TOBG2	47	319.2
TOBG3	41	338.4
RFP1	41	77.7
RFP2	41	32.7
RFP3	42	59

Glycerol stocks

• Took 750ul from overnight culture and centrifuged at full speed for ~2 minutes
• Supernatant was removed
• Pellet resuspended in 375ul of HMFM
• Samples were then frozen at -80

Gel Purification of the pGEM-T pasmid double digestions ran in the 11/09/13

The ge was purified by using the The Zymoclean Gel DNA recovery Kit and its protocol was followed.

Nanodrop Results

Sample	Volume	Concentration (ng/ul)	280/260	260/230
TODX	48.5	7.1	1.83	0.72
TODF	45.5	21.7	1.89	1.19
TOBG	47.7	9	1.77	0.70
pSB1C3 (RFP)	46.2	7.3	1.81	0.16

Digestion of the samples purified with the enzyme PstI-HF

DNA volume	Volume	Cutsmart buffer	Enzyme volume	dH2O added to total volume of 60 ul
TODX	23	6	1	30
TODF	22.5	6	1	30.5
TOBG	22	6	1	31
pSB1C3 (RFP)	20.5	6	1	32.5