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August										September
Mon	Tue	Wed	Thr	Fri	Sat	Sun				Mon	Tue	Wed	Thr	Fri	Sat	Sun
				1	2	3				1	2	3	4	5	6	7
4	5	6	7	8	9	10				8	9	10	11	12	13	14
11	12	13	14	15	16	17				15	16	17	18	19	20	21
18	19	20	21	22	23	24				22	23	24	25	26	27	28
25	26	27	28	29	30	31				29	30

August

August 11th

Restriction Digests

object	restriction enzymes
His-RFP with stop codon	SpeI,EcoRI
5'-intron M	SpeI,EcoRI
Lac-3'intron-RBS	SpeI,EcoRI
His-RFP without stop codon	XbaI,EcoRI
RBS-Amp	SpeI,EcoRI

Ligation

• EcoRI-His-RFP(with stop codon)-SpeI + XbaI-DT-Cm-EcoRI
• EcoRI-5'intron M-SpeI + XbaI-DT-Cm-EcoRI
• EcoRI-Lac-3'intron-RBS-SpeI + XbaI-His-RFP(without stop codon)-5'intron-DT-Cm-EcoRI
• EcoRI-His-RFP(without stop codon)-SpeI + SpeI-Amp-EcoRI

Transformation

• His-RFP(with stop codon)-DT-Cm
• 5'intron M-DT-Cm
• Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT-Cm
• His-RFP(without stop codon)-Amp

Nano drop

object	concentration(ng/µL)
His-RFP(with stop codon) EcoRI-SpeI	162.6
5'intron M EcoRI-SpeI	45.4
Lac-3'intron-RBS EcoRI-SpeI	120.9
His-RFP(without stop codon) EcoRI-SpeI	160.0
His-RFP(without stop codon)-5'intron-DT-Cm XbaI-EcoRI	174.0
Amp EcoRI-SpeI	101.8
Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT

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August 12th

Colony PCR

Electrophoresis

Shaking culture

To analysis sequence

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August 13th

sasisuseso

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