Revision as of 02:04, 1 September 2014 by Fukufuku (Talk | contribs)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

>

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 Enzyme Digestion

sample	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

Densitometry

sample	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

back to calender

August 12th

Colony PCR

• 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:[1]turning red
• Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT-Cm:[2]turning pink
• Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT-Cm:[3]turning white
• His-RFP(without stop codon)-Amp

Electrophoresis

1. His-RFP(with stop codon)-DT
2. His-RFP
3. 5'intron M-DT
4. 5'intron M
5. Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT:[1]
6. Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT:[2]
7. Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT:[3]
8. His-RFP(without stop codon)-5'intron-DT
9. His-RFP(without stop codon)

Liquid Culture

• His-RFP(with stop codon)-DT
• 5'intron M-DT
• Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT:[1]
• Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT:[3]
• His-RFP(without stop codon)

back to calender

August 13th

Miniprep

PCR

By use of KAPA HiFi DNA Polymerase
• His-RFP(with stop codon)-DT-Cm
• 5'intron M-DT-Cm
By use of Taq DNA Polymerase
• Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT-Cm:[1]turning red
• Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT-Cm:[3]turning white

Electrophoresis

Restriction Enzyme Digestion

Ligation

Transformation

back to calender

August 16th

Liquid Culture

back to calender

August 18th

Miniprep

Colony PCR

Densitometry

Ligation

Transformation

Analysis of DNA sequences

back to calender

August 19th

Colony PCR

Electrophoresis

Shaking Culture

back to calender

August 20th

Colony PCR

Same as August 19th Colony PCR.

back to calender

August 21th

Reagent Preparation

Protein Extraction

SDS-PAGE

back to calender

August 22th

Prorein Purification by use of Ni-NTA

SDS-PAGE

back to calender

August 23th

Prorein Purification by use of Ni-NTA

SDS-PAGE

back to calender

August 24th

Extraction of plasmid device

back to calender

August 25th

Extraction of plasmid device

SDS-PAGE

back to calender

August 26th

PCR

Restriction Enzyme Digestion

Densitometry

Ligation

Transformation

back to calender

August 27th

Colony PCR

Shaking Culture

back to calender

August 28th

back to calender