Team:Gifu/Notebook
From 2014.igem.org
(Difference between revisions)
Line 159: | Line 159: | ||
<h3>PCR</h3> | <h3>PCR</h3> | ||
<ul class="list"> | <ul class="list"> | ||
- | <ul>By use of KAPA</ul> | + | <ul>By use of KAPA HiFi DNA Polymerase</ul> |
+ | <li>His-RFP(with stop codon)-DT-Cm</li> | ||
+ | <li>5'intron M-DT-Cm</li> | ||
+ | <ul>By use of Taq DNA Polymerase</ui> | ||
+ | <li> | ||
<h3>Electrophoresis</h3> | <h3>Electrophoresis</h3> | ||
<h3>Restriction Enzyme Digestion</h3> | <h3>Restriction Enzyme Digestion</h3> |
Revision as of 08:10, 30 August 2014
Calender
Click on a date to see our research activities on that day.
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 |
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
- His-RFP(without stop codon)-Amp
Electrophoresis
- His-RFP(with stop codon)-DT
- His-RFP
- 5'intron M-DT
- 5'intron M
- 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:[2]
- Lac-3'intron-RBS-His-RFP(without stop codon)-5'intron-DT:[3]
- His-RFP(without stop codon)-5'intron-DT
- 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)
- His-RFP(with stop codon)-DT-Cm
- 5'intron M-DT-Cm
-
Electrophoresis
Restriction Enzyme Digestion
Ligation
Transformation
August 16th
Liquid Culture
August 18th
Miniprep
Colony PCR
Densitometry
Ligation
Transformation
Analysis of DNA sequences
August 19th
Colony PCR
Electrophoresis
Shaking Culture
August 20th
Colony PCR
Same as August 19th Colony PCR.
August 21th
Reagent Preparation
Protein Extraction
SDS-PAGE
August 22th
Prorein Purification by use of Ni-NTA
SDS-PAGE
August 23th
Prorein Purification by use of Ni-NTA
SDS-PAGE
August 24th
Extraction of plasmid device
August 25th
Extraction of plasmid device
SDS-PAGE
August 26th
PCR
Restriction Enzyme Digestion
Densitometry
Ligation
Transformation
August 27th
Colony PCR
Shaking Culture
August 28th
August 13th
Miniprep
PCR
- By use of KAPA HiFi DNA Polymerase
- By use of Taq DNA Polymerase