Team:UT-Tokyo/Notebook
From 2012.igem.org
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== Lab Notes == | == Lab Notes == |
Revision as of 01:44, 27 September 2012
Notebook
![box-background image](https://static.igem.org/mediawiki/2012/8/8a/UT-Tokyo_LN-02.jpg)
Correspondence table
No. | Biobrick Parts |
---|---|
#1 | pConst |
#2 | Argbox |
#3 | RBS |
#4 | Argbox×8 |
#5 | d.term |
#6 | fake of fhlA |
#7 | fhlA(P-siteless) |
#8 | hycAp |
#9 | Ptrc-RBS-rocF |
#10 | fhlA(E363G) |
#11 | RFP(psb1K3) |
#20 | pLac |
#40 | RBS(weak)-GFP-d.term |
#44 | RBS(strong)-GFP-d.term |
#66 | RFP(psb1C3) |
Lab Notes
May
1st week: 5/27-6/2
May 29th
- Making LB medium
May 30th
- Making LB Agar
- Making LB Amp Plates
May 31st
- Preculture
June
2nd week: 6/3-6/9
June 5th
- Plating JM109
June 6th
- preculture
June 7th
- Measuring OD by spectroscope
- Meking electrocompetent cells
- Electroporating of a test plasmid
June 8th
- Caluculatiing transformation efficiency
3rd week: 6/10-6/16
none
4th week: 6/17-6/23
none
5th week: 6/24-6/30
June 26th
- Making LB tetracycline plate
June 27th
June 28th
- Making LB kanamycin plate and chloramphenicol plate
- Transformation with a test plasmid
June 29th
- Transformation with #1(constitutive promoter) and #11(RBS)
July
6th week: 7/1-7/7
July 3rd
- Making K solution
- Colony PCR DNA fragment assumed as fhlA (#6) for cloning
July 4th
- Miniprep #1, #11
July 5th
- Making 1% agarose solution
- Electrophoresis (#6)
- Digest (#1_EP, #11-_EP)
July 6th
- Colony PCR (#6)
7th week: 7/8-7/14
July 9th
- Electrophoresis (#6)
July 10th
- Transformation
- #3(RBS medium), #5(d.term), #20(pLac)
July 11th
- Preculture
- #3, #5, #20
- Gel Extraction(#6)
July 12th
- The continuation of Gel Extraction(#6)
- Miniprep
- #3, #5, #20
July 13th
- Nanodrop (#6).
- Digestion (#3:E-P, #5:E-P, #20:E-P)
- Electrophoresis (#3, #5, #20)
July 14th
- Miniprep (#1,3,5,11,20)
- Gel extraction (#6)
8th week: 7/15-7/21
July 17th
- Transformation (#1A2,#3,#5,#20)
- PCR (parts of 1. and #3,#20(Colony PCR))
- Incubate #5.
July 18th
- Miniprep (#1A2,#3,#5,#20)
- PCR using u.p (#1A2,#3,#5,#20)
- Colony PCR (#3,#20,H₂O)
- Electrophoresis (#1A2,#3,#5,#20, #3,#20, H₂O)
- PCR (2011’s #1,#36 and 2012’s #3,#5,#20, H₂O)
July 20th
- Miniprep.
- Electrophoresis (7/19’s PCR product(#1,#3,#5,#20,#36) )
- Colony PCR (#3,#5,#20)
July 21st
- Electrophoresis (7/20’s PCR product (#3,#5,#20))
9th week: 7/22-7/28
July 24th
- Making agar medium plate contain ampicillin
- PCR (#6, #8)
- Electrophoresis (#6, #8)
- Transformation (#1)
- Culture (#5, #20)
July 25th
- Miniprep (#5, #20)
- Purify (#6, #8)
- Digestion (#3:S-P,#5:E-X,#11:E-S,#20:S-P)
- Electrophoresis (#5, #11)
- Ligation (#5,#11) (?)
July 26th
- Culture (#1)
- Digestion (#3:S-P, #5:E-X, #11:E-S, #20:S-P, fhlA:E-S, hycA:E-S)
- Electrophoresis (#3:S-P, #5:E-X, #11:E-S, #20:S-P, fhlA:E-S, hycA:E-S)
- Gel extraction (#3:S-P, #5:E-X, #11:E-S, #20:S-P, fhlA:E-S, hycA:E-S)
July 27th
- Culture, Miniprep (#1)
- Digestion (#5-A:E-X, #11-A:X-P)
- Electrophoresis (#3:S-P, #20:S-P, #11:E-S)
- Gel extraction (#3:S-P, #20:S-P, #11:E-S, E-S vector, hycAp:E-S)
July 28th
- Digestion(#1,3,5,11,hycAp)
- Gel extraction(#1,3,5,11,hycAp)
- Cloning(fhlA)
10th week: 7/29-8/4
July 30th
- Digestion (#3:S-P, #11:X-P)
- Electrophoresis (#3, #11)
- Gel extraction (#3, #11)
- Ligation (#3, #11)
- Transformation (ligation product; #3-11)
August 1st
- Digestion (#3:S-P, #11:X-P)
- Electrophoresis (#3, #11)
- Gel extraction (#3, #11)
- Ligation (#3, #11)
- Transformation (ligation product; #3-11)
- Miniprep (#3, #5, #11)
August 2nd
- Colony PCR (#3-11)
- Culture (#3-11)
- Colony pick up, culture (red colony of #11)
- Digestion (#3:S-P, #5:E-X)
August 3th
- Gel extraction (#3:S-P, #5:E-X)
- Making solution of Kanamycin (100mg/mL)
- Culture, miniprep (#11)
- Culture (#11)
August 4th
- Plating, Miniprep (#11)
August
11th week: 8/5-8/11
August 6th
- Culture, Colony PCR (#3-11)
August 7th
- Colony PCR, culture (#3-11)
- Miniprep, Digestion, Gel extraction (culture product; #3-11)
August 8th
- Digestion (#5: E-X)
- Gel extraction (#5)
- Miniprep (#5)
- Colony PCR (culture product; #3-11)
August 9th
- Colony PCR (#3-11)
- Culture, Miniprep (#11)
- Digestion (#11:X-P)
- Cloning (#6,#8)
- Ligation
- Gel extraction (ligation product; #5, #11)
- Ligation, Transformation (#3, #11)
August 10th
- Colony PCR (#3-11)
- Miniprep (#3)
- Digestion (#3:S-P)
- Cloning, Colony PCR (#6,#8)
- cut check, PCR check (#3-11)
- Electrophoresis (ligation product; #3, #11, PCR product; #6)
- Gel extraction (#6)
August 11th
- electrophoresis(PCR product; #3-11)
- Digestion (#6:E-S)
- electrophoresis (#8)
12th week: 8/12-8/18
August 13th
- Check, Culture (#3-11)
- Transformation (test plasmid)
August 15th
- Gel extraction (cut check product; #3, #11)
- Ligation, Transformation (#3-#11)
- Make Vector for #6
- Digestion (#3: E-S)
- Gel extraction (digestion product; #3, #6(8/11)
- PCR check (ligation product; #3-11)
- Cloning (#6)
August 16th
- Gel extraction (#6)
- cloning (#6)
- PCR check (#3-11)
- PCR-cleanup (#8)
- culture (#3-11)
August 17th
- electrophoresis (#6)
- Miniprep (#3-11)
- Digestion (#3-#11: E-S, #6: E-S)
- Gel extraction (#3-11)
- Ligation, Transformation (#3-11-5)
- Making solution of Ampicillin
13th week: 8/19-8/25
August 20th
- PCR check (#3-11-5)
- Digestion (#8: E-S)
- Cloning (#6)
- Digeston (#8: E-S)
- Gel extraction (#6)
August 21th
- Colony PCR (#1)
- Digestion (#6: E-S, #3-11: E-S, #20: S-P)
- Ligation (#3-11, #5)
- Gel extraction (#3-11, #6)
- Ligation (#6-pSB1A2)
- Electrophresis (#3, #5; made previously)
August 22th
- Cloning (#6; from test plasmid)
- Colony PCR (#3-11-5(colony is not red), #20, 8/21 ligation product;#6, #8)
- Miniprep,Frozen stock (culture product; #1, #5, #20)
- Transformation (#8, #3-11-5)
- Digestion (#6:X-S)
- Making solution of Ampicillin
- cloning retry (#6)
August 23th
- PCR check (ligation product; #6)
- Cloning (#6)
Colony PCR (#1, #6, #8, #11, #3-11-5, #3-11)
August 24th
- Digestion (#6, E-S, #3-11, E-S)
- Colony PCR #3-11
- Electrophresis, Gel extraction (#6)
- Vector insert (#6)
- Sequencing (#8)
August 25th
- Digestion (#6: E-S)
- Colony PCR (#6-Vector)
- Gel extraction (#3-11)
14th week: 8/26-9/2
August 26th
- Electrophresis (#6)
- Master plate (#6)
- Colony PCR (#3-11-5)
- Preculture (#6)
August 27th
- Ligation (Miniprep product; #3-11, #5)
- Digestion retry (#3-11, #5)
- Miniprep (#6)
- Mutagenesis (#6)
- Sequencing (#6)
August 28th
- Mutagenesis(#6)
- Digestion(#3-11,#5)
August 29th
- Mutagenesis by inverse PCR(#6)
- Digestion(#5)
- Cut check(#6)
- PCR check(#6)
- Sequencing(#6)
- Making agar medium plate contain Chloramphenicol
August 30th
- Digestion(#5,#3-11,#8)
- Sequencing(#6)
- Mutagenesis(#6)
August 31st
- Colony PCR(#6)
- Digestion(#3-11,#5)
- Ligation(#3-11+#5)
- Transformation(#3-11-5,#66)
- Culture(#66)
September 1st
- Cloning(#6)
- Miniprep(#66)
September
15th week: 9/3-9/9
September 3rd
- Miniprep(#3,#11)
- Colony PCR(#3-11-5)
- Transformation(#2)
September 4th
- Digestion(#3,#6,#11)
- Ligation(#3,#6)
- Miniprep(#2)
September 5th
- Ligation(#6+1A2)
- Check PCR(#6)
- Digestion(#2,8)
September 6th
- Mutagenesis(#7)
- Sequencing(#6)
- Digestion(#5,#66)
- Transformation(#2,#6)
September 7th
- Colony PCR(#2,#8)
- Cut check(EX,ES,XP,SPcut)
- Digestion(#5,#66)
- Ligation(#66+#5)
- Transformation(#66-5)
- Mutagenesis(#7)
September 8th
- Sequencing(#6)
- Colony PCR(#66-5)
- Miniprep(#2)
16th week: 9/10-9/16
September 10th
- Mutagenesis(#7)
- Digestion(#2)
- Transformation(#9,#40)
September 11th
- Transformation(#44)
- Mutagenesis(#7)
- Miniprep(#6)
- Colony PCR(#7)
September 12th
- Culture(#44)
- Miniprep(#44)
- Mutagenesis(#7)
- Transformation(#7)
September 13th
- Culture(#1,#7,#20)
- Colony PCR(#7)
- Mutagenesis(#7)
September 14th
- Inverse PCR(#10)
- Digestion(#3,#7)
- Ligation(#3-7)
- Transformation(#3-7,#10)
September 15th
- Digestion(#1,#7,#8,#44,#66)
- Sequencing(#7,#10)
- Colony PCR(#44,#3-7)
September 16th
- Digestion(#1,#3,#7-5,#8,#10,#44,#66)
- Transformation(#3-7-5,#3-10)
17th week: 9/17-9/23
September 17th
- Miniprep(#7-5,#44)
- Digetion(#3,5,10)
- Transformation(#3-7-5,3-10-5)
September 18th
- Miniprep(#7-5,#44)
- Digetion(#3,#5,#10)
- Transformation(#3-7-5,#3-10-5)
September 19th
- Transformation(#4)
- Miniprep(#3-7-5,#3-10-5)
- Digestion(#3-7-5,#3-10-5,#20,#66)
September 20th
- Miniprep(#4)
- Digestion(#3-7-5,#3-10-5,#4,#20,#66)
- Transformation(#20-3-7-5,#20-3-10-5,#4-4)
September 21st
- Digestion(#3-7-5,#3-10-5,#4,#20,#66)
- Transformation(#20-3-7-5,#20-3-10-5,#4-4,#7+1C3,#10+1C3)
September 22nd
- Digestion(#3-7-5,#3-10-5,#4,#20,#66)
- Transformation(#20-3-7-5,#20-3-10-5,#4-4)
- Miniprep(#7+1C3,#10+1C3)
September 23rd
- Digestion(#3-7-5,#3-10-5,#4,#8,#20,#66)
- Transformation(#20-3-7-5,#20-3-10-5,#4-4)
- Colony PCR(#20-3-7-5)
18th week: 9/24-9/26
September 24th
- Digestion(#3-7-5,#3-10-5,#4,#8,#20,#66)
- Transformation(#20-3-7-5,#20-3-10-5,#4-4)
- Colony PCR(#20-3-7-5,#4+1C3)
September 25th
- Digestion(#3-7-5,#3-10-5,#4,#20,#66)
- Transformation(#20-3-7-5,#20-3-10-5,#4-4)
- Colony PCR(#8-44)
September 26th
- Digestion(#20-3-7-5,#20-3-10-5,#4,#66,#8-44)
- Transformation(#8-44-20-3-7-5-1C3,#8-44-20-3-10-5-1C3)
- Culture(4-4-1C3)