Team:UT-Tokyo/LabNote

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Revision as of 17:16, 2 October 2011 by Hiro3726 (Talk | contribs)

Parts List

Each part is represented by s number (e.g. #20 for lac promotor) in this notebook. We have also used this number in our hand-written lab notebook. By pointing the mouse on the number in the construct, you can find out the details of the part.


Table 1. List of iGEM parts used in our project
Number Reg.ID Content Plasmid Length (bp)
#1 J23119 c.Promoter (Strong) pSB1A2 35
#2 J23118 c.Promoter (Medium) BBa_J61002 35
#3 B0032 RBS pSB1A2 13
#5 B0014 d.Ter pSB1AK3 95
#9 E0240 RBS-GFP-d.Ter pSB1A2 876
#10 E0040 GFP pSB1A2 681
#11 E1010 RFP pSB2K3 723
#14 I712019 fLuc pSB1AK8 1653
#17 J52008 rLuc pSB1AK3 936
#20 R0011 lacP pSB1A2 55
#21 C0012 LacI pSB1A2 1128
#22 I712074 pT7 pSB1AK8 46
#23 K145001 T7 RNA Pol. pSB1A2 2655
#24 J22106 recAp pSB1A2 192
#27 C0083 AspA pSB2K3 1518
#28 K112808 T4 phage lysis device (no promoter) pSB1A2 1785
#29 - CheZ pSB1AK3 728
#30 K117000 Lysis gene pSB1A2 144
#31 - LexA pSB1AK3 750
#33 - sulAp pSB1AK3 67
#34 - uvrAp pSB1AK3 96
#35 - recNp
#36 R0051 cI-repressed promoter pSB1A2 49
#37 C0051 cI repressor (LVA tagged) pSB1A2 \
#38 - RecA

Lab Diary

  • May
  • June
  • July
  • August
  • September
  • October

Please enable Javascript to view this calendar.

May

'11/05/18 (Wed)

  • Making LB medium, 50×TAE, Tris-HCl (pH8.0)

'11/05/24 (Tue)

  • Making SOB medium, 0.5M EDTA (pH8.0)

'11/05/25(Wed)

  • Making TB(pH=6.7), LB plate

'11/05/26(Thu)

  • Making Mgaq(for SOB medium), Competent cell
  • TB filtration

'11/05/31(Tue)

  • iGEM parts resuspension + frozen stock (at -20℃)
  • Transformation
  • Overnight culture on LB plate (with 100ug/ml ampicilline)

June

'11/06/01(Wed)

  • Picking up colony and transfer to LB medium
  • Making LB medium

'11/06/02(Thu)

  • Miniprep
  • Making Glycerol(50%)(for cryopreservation)

'11/06/07(Tue)

  • Nanodrop
  • Transformation
  • Culture from frozen stock

'11/06/08(Wed)

  • Picking up colony

'11/06/09(Thu)

  • Miniprep

'11/06/13(Mon)

  • Planting Negative control
  • Making frozen stock
  • Transformation(BBa_E0240, #3#5#10

'11/06/14(Tue)

  • Picking up colony
  • Making Mg reagent

'11/06/15(Wed)

  • Miniprep

'11/06/17(Fri)

  • Picking up colony(#3#9#5#1#2#24

'11/06/21(Tue)

  • Making frozen stock(#3#9#9

'11/06/22(Wed)

  • Miniprep(#9

'11/06/24(Fri)

  • Digest (product of Miniprep 06/09 EScut)
  • Making agarose gel
  • Electrophoresis

'11/06/28(Tue)

  • Digest(product of Miniprep 06/09 EPcut)
  • Electrophoresis
  • Defrost and transformation(BBa_E0030, E0020, #14#17

'11/06/29(Wed)

  • Picking up colony
  • Making LB broth

July

'11/07/01(Fri)

  • Digest(EPcut)
  • Miniprep(BBa_E0030, E0020, #14#17

'11/07/05(Tue)

  • Electrophoresis(product of digest 07/01)
  • Digest(#9#14#17#3#5

'11/07/06(Wed)

  • Gel extraction (pre)(BBa_I712052)
  • Picking up colony(Ba_J23119, #2#24#10

'11/07/07(Thu)

  • Digest（BBa_E0240, #14#17#3#5#20#21#22#23#20#21#22

'11/07/08(Fri)

  • Miniprep(BBa_J23119, #2#10#24

'11/07/11(Mon)

  • Gel extraction(#9#14#17#1#11#20#21#22#23

'11/07/12(Tue)

  • Picking up colony

'11/07/13(Wed)

  • Frozen stock(#1#20#21#14#23#11#3#5#2

'11/07/14(Thu)

  • Miniprep(#1#11#20#21#22#23#2#2#3#5#11

'11/07/15(Fri)

  • Ligation(#2-#9#2-#3#14-#5#17-#5#2-#9#2-#3#14-#5#17-#5#11

'11/07/19(Tue)

  • Digest
  • Gel extraction
  • Making competent cell

'11/07/20(Wed)

  • Picking up colony
  • Making Master plate
  • Testing product of Miniprep
  • Transformation (#2#27#28

'11/07/21(Thu)

  • Miniprep(#2-3#2-9#14-5#17-5#11#20#20#2-3#10#11#14-5#17-5

'11/07/22(Fri)

  • Frozen stock(#2#2-3#2-9#14-5#17-5#20#2-3#10#11#14-5#17-5#20#10#11#14-5#17-5#27#28

'11/07/25(Mon)

  • Colony check(#2#2-9#20#10#11#14-5#17-5#20-#9#3-#11#3-#14-5#3-#17-5

'11/07/26(Tue)

  • Picking up colony and making master plate(#20-9#3-11#3-14-5#3-17-5#20-9#3-11#3-14-5#3-17-5#14-5#22#23#3-#14-5#3#15#27#28#30#3-14-5#3

'11/07/27(Wed)

  • Miniprep(#1#2#3#10#22#24#20-9#3-11#3-17-5

'11/07/28(Thu)

  • Gel extraction(#14-5#22#23#1#2#3#24#3-11#3-17-5#3-#14-5

'11/07/29(Fri)

  • Cloning(lexA, cheZ)
  • Colony PCR(#3-#14-5#1#2#3#24#3-11#3-17-5#30

August

'11/08/01(Mon)

  • Nanodrop(#3-11#1#3#24#2#3-17-5#30#3#5#9#30#3#9#30#2-#3-17-5#3-11-#5#14-#5#3-#23#22-#9#3-#30#2-#3-17-5#3-11-#5#14-#5#3-#23#22-#9#3-#30#27#28

'11/08/02(Tue)

  • Colony PCR(#2-#3-17-5#3-11-#5#14-#5#3-#23#22-#9#27#28#27#28#30

'11/08/04(Thu)

  • Electrophoresis(product of ligation #2-3-17-5#3-11-5#14-5#22-9#2-3-17-5#3-11-5#14-5#22-9#23#20-9

'11/08/05(Fri)

  • Digest(#14-5#3-11-5#27#28

'11/08/08(Mon)

  • Defrost Frozen stock, Miniprep, Nanodrop and Electrophoresis(#5#20#23#30#27#28#3-11-5#23#30#6#7#7#20#5#14-5#3-11-5

'11/08/09(Tue)

  • Miniprep, Electrophoresis and Digest(#5#14-5#3-11-5#6#27#8#3-11-5#23#7#20#7#27-#3-27#20-28#28-#3-29#29-#31-#3-#23#1-#3-11-5#2-#3-11-5#24-#3-11-5#3-#30

'11/08/10(Wed)

  • Colony PCR(#27-#3-27#20-28#28-#3-29#29-#31-#3-#23#1-#3-11-5#2-#3-11-5#24-#3-11-5#3-#30#6#7#1#2#3#7#20#23

'11/08/11(Thu)

  • Gel extraction(#30#14-5#5#3-11-5#3#20-28#31#24-3-11-5#3-30

'11/08/12(Fri)

  • Colony PCR(#3-14-5#6-5#3-23#27#28#3-14-5#5#5#14#5#14-5#3-30#27#28#13-14-5#6-5#3-23#27#28#6-5#3-23

'11/08/15(Mon)

  • Gel extraction(from previous digest products(8/12) : #27#28#33#34#4#3-23#3-11-5#3-11-5#3-#14-5#24-#3-17-5#24-#3-11-5#20-#28#3-#27#3-#29#3-23-#5#33-#34--#35--#2-9#14#17 37 incubation

'11/08/16(Tue)

  • Colony PCR(#3-#14-5#24-#3-17-5#20-#28#3-#27#3-#29#3-23-#5#20-#28#5#14#17#2-9#2-4#2-3-17-5#1#2#24-#3-11-5#33-#34-#35-#3#3#3-#14-5#3-#27#3-#29#3#14#17#14#2-3-17-5#14#17#1#2#3

'11/08/17(Wed)

  • Hotstart-PCR (Change conditions a little)
    • From Genome
    • From nested-region
    • From parts
  • Making Competent Cell, SOB medium
  • Ethanol precipitation(WT: 40ng/ul)
  • Culture from master-plate(#6-5#20-28#3-27#33#24-3-11-5#24-3-17-5 Miniprep *Assay(IPTG induced cell lysis device *Gel extraction(SPcut:#3#1#2#2-3-17-5#3#6-5#20-28#3-27#33#24-3-11-5#24-3-17-5

'11/08/18(Thu)

  • Digest(8/17#3#33#14-5#33#3-27#3-30#20-28#2-3-11-5#3#33#14-5#33#3-27#1#2#2-3-11-5#2-3-17-5#1#2#2-3-17-5#2-3-11-5#3-30#20-28#3-#14-5#3-8#29#33-#3-11-5#34-#3-27-#5#20-28-#28-

'11/08/19(Fri)

  • ColonyPCR(from ligation products)(#3-14-5#3-29#33-3-11-5#34-#3-27-5#1#2#2-3-11-5#2-3-17-5#2-3-11-5#3-29#34-#3#3-27#3-30#6-5#7#20-28#24-3-11-5#24-3-17-5#33

'11/08/22(Mon)

  • Making SOB medium
  • Culture for Miniprep
    • From frozen stock(#1#2#7#2-3-17-5#3-30#3-14-5#33-3-11-5#34-#3-27-5#1#2#2-3-17-5#3-30#7#7#3-14-5#3-27-5#34#33-3-11-5#7#30#2-3-17-5#3-30#3-14-5#34-#3-27-5#33#24-3-17-5#14-5

'11/08/23(Tue)

  • Gel extraction
    • from O.N. digest products(#7#30#2-3-17-5#3-30#3-14-5#34-#3-27-5#24-3-17-5#14-5#34#3-27-5#33-3-11-5#3-14-5#33-3-11-5#34-#3-27-5#14-5#3-27-5#3-27-5#14-#5#3-30-#5#3-#30#34-#3-11-5#20-#28#33

'11/08/24(Wed)

  • Gel extraction & Electrophoresis(#3-27-5#14-5#3-27-5 PCR clean-up system -> digest **#27 XP cut **#28 XP cut *Making plate and amp stock　->4℃ **amp 32 **km 11 (km conc. = 3/4 * that it was) *Miniprep and Nanodrop **#33transfromation *Luciferase assay reagents prep **D-Luciferin Solution : 1ml*10 + 100ul*10 at -80℃ **Coelenterazine Solution : 100ul * 10 at -20℃ **Lysis buffer : 500ul * 10 at -20℃ *Ligation(#3-30-#5#3-11-5-#34#30-#3#28-#20#35-#2-3-11-5-#14-5-#3

'11/08/25(Thu)

  • Renilla luciferase assay (with #2-3-17-5#27#28#20-#28#3-#27#14-5#33#14#2-3-17-5

'11/08/26(Fri)

  • Renilla luciferase assay (with #2-3-17-5#33#14#5#2-3-17-5

'11/08/27(Sat)

  • Ligation & Transformation (0826 ①②③⑤⑧⑨) (into Nippon Gene comp.)
  • Contamination check (WT FS, ΔcheZ Frozen stock)
  • Waking from Frozen stock(WT, ΔcheZ)

'11/08/28(Sun)

  • Making ΔcheZ comp., LB amp plates, TB
  • Colony PCR & master plate preparation -> Failure!!
  • Gel extraction (#29#5#33#14-5#20#14-5#20#14-5

'11/08/29(Mon)

  • PCR purification -> digest with XP(#27#28#20#14-5#14-5#20#3-#14-5#2-3-11-5#35-#33#34#3-17-5

'11/08/30(Tue)

  • Making LB amp, Aspartic acid solution (10mM), H2O2 solution (10mM)
  • Gel extraction -> PCR was failed!(#27#28#3-#14-5#2-3-11-5#35-#33#34#3-17-5 check (5uL EP -> PCR clean-up(#27#28#20#3#14-5#20#3-#29#2-3-11-5#3#2-9 Line-drawing plating *SOS stress (UV irradiation or H2O2)(#24-3-11-5#33-3-11-5#34-3-17-5#24-3-11-5#33-3-11-5

'11/08/31(Wed)

  • Making LB plates(amp, km)
  • Luciferin reagent preparation
  • Flask A.C. for making competent cells
  • Miniprep (#34-3-17-5#20#3 clean-up -> XPcut(#27#28#14-5#3-30#2-9#24-3-17-5#2-3-17-5#20-#3-17-5#2-3-11-5-#3-#14#20-#28#3-#27#3-#29#35-#3-#14-5#3-30-#5#33-#3-17-5#34-3-17-5#36#37#3-17-5#2-3-11-5#2-3-11-5

September

'11/09/01(Thu)

  • Colony PCR
    • #34-3-17-5#3-14#3-29#35-#3-14-5#37#27#28#27#28#3-30 failed!, #14-5#3#20#3-14#3-29#3-14-5#3 Failed (Mixed with DMSO!), #20#3-14#3-29#3-14-5#3-27#20-#3-17-5#36#3#20#14-5

'11/09/02(Fri)

  • Making medium and reagent(LB 120ml(0.125, 0.25, 0.5% Agar, M9 stock(20×), M9 200ml, Vitamins Solution (100×))
  • Gel extraction -> Digest products were OK but extraction was failed.
    • #20#3-14#3-14-5#3-29#20-3-17-5#3-27#3#20#37#3-14#3-29#3-14-5#20-3-17-5#3-27 PCR products were OK but all purified products were lost. **#20-3-17-5#3-27#3-14#3-14-5#3-29#20-3-17-5#3-27#3-14-5#3-29#3#20#37#3-14#3-29#3-14-5#20-3-17-5#3-27#3-14-5#3-29

'11/09/03(Sat)

  • Gel extraction
  • Ligation & Transformation
    • #20-#3-14-5#2-#3-14-5#3-27-#5#3-27-#5#37-#5#36

'11/09/04(Sun)

  • Hot-start PCR Pre-Mix (4×) preparation (Buffer + dNTPs + THUNDER Taq)
  • Colony PCR(#20-3-14-5#3-27-5#3-29-5#37-5#2-#3-14-5#3-27-#5#3-29-#5#37-#5#20-#28#36#2#5#20-3-14-5#20#20

'11/09/05(Mon)

  • Colony Diffusion Assays (#2-9#20#20-3-14-5#20-3-17-5#2-#3-14-5#3-27-#5#3-29-#5#37-#5#20-#28#36 clean-up -> Digest(#3-29-#5#37-5#20-3-14-5#37-5#36 Asp -> all failed(.125%, .25% Agar) *Culture for Miniprep(#2-3-14-5

'11/09/06(Tue)

  • Gel extraction(#37-5#3-29-5#2-3-14-5#20-3-14-5#20-3-17-5#36#3-29-5#37-5 Purification -> Digest(#3-29-5#37-5#3-27-#5#3-30-#5#2-3-11-5-#5#2-9 4℃ storage *Ninhydrin sol. preparation (Not dissolvable!) -> 4℃ storage

'11/09/07(Wed)

  • Colony PCR(#3-27-5#3-30-5#36#36#3-29-5#37-5#36#3-29-5#37-5#3-27-5#3-29-5#37-5#36 Digest(#3-14-5#3-29-5#37-5#3-27-5#36#2#2-3-17-5#3-14-5#34#2#34#36#3-27-5#3-14-5#2-3-17-5#33-#3-14-5#34-#3-14-5#36-#3-14-5#2-#3-29-5#20-#3-29-5#3-#37-5#2-3-11-5-#1-#3-17-5#33#34#36#1#2#20#3

'11/09/08(Thu)

  • Making competent cell (WT), LB(amp) plate
  • Frozen stocks(#36#3-14-5#3-27-5#33-3-14-5#34-3-14-5#36-3-14-5#20-3-29-5#2-3-11-5#2#34#36#2-3-17-5#3-27-5#3-14-5#35#31#3-14-5#3-27-5#3-29-5#37-5#2#34#36#2-3-17-5#3-14-5#34-3-14-5#36-3-14-5#20-3-29-5#20-3-14-5 Store at -20C(#33-3-11-5#34-3-14-5#20-3-29-5#37-5#35#31#31#3-#37-5#36-3-29-5#3-14-5#2-3-17-5#20-#3-27-5#34-3-17-5#36-3-14-5#20-3-29-5#2-3-11-5#34-3-14-5#36-3-14-5#20-3-14-5

'11/09/09(Fri)

  • Making competent cell (WT), TB
  • Colony PCR(#35-#31-#3-#37-5#36-#3-29-5#3-14-5-#2-3-17-5#20-3-27-5#3-14-5#3-17-5#31#3-37-5#36-3-29-5#3-14-5-2-3-17-5#3-11-5#3-14-5#3-17-5#31#3-37-5#36-3-29-5#34-3-14-5#36-3-14-5#20-3-29-5#2-3-11-5#34-3-14-5#36-3-14-5#20-3-14-5#3-11-5#3-11-5#29#36-3-14-5#2-3-17-5#20#3-27-5#2-3-11-5#20-3-29-5#20-3-29-5#31-#3-37-5#36-3-29-5#3-14-5-2-3-17-5 Luciferase Assay(#24-3-17-5#34-3-17-5 Sequencing(#33-3-11-5#34-3-14-5#20-3-29-5#37-5

'11/09/10(Sat)

  • Culture plate storage (4℃)
  • Miniprep(#31-#3-37-5#36-3-29-5#3-14-5-2-3-17-5#3-14-5#3-17-5#31#3-37-5#36-3-29-5#36-3-14-5-#2-3-17-5#20#3-37-5

'11/09/11(Sun)

  • Digest from Miniprep products
    • #3-14-5-2-3-17-5#36-3-14-5#20 O.N. *Colony PCR **#3-11-5#29#20-#3-27-5#20-#3-37-5#36-#3-14-5-2-3-17-5#3-14-5-2-3-17-5#20-3-37-5#29#20#2-3-17-5#36-3-14-5-2-3-17-5#20#3-14-5-2-3-17-5#1#3-14-5-2-3-17-5 Digest for >1h -> Heat kill for 20min (#20-3-37-5#36-3-14-5-2-3-17-51h -> Heat kill for 20min (#36-3-14-5-2-3-17-5#2-3-17-5#20-3-14-5#2-3-17-5#36-3-14-5-2-3-17-5#20-3-37-5#36-3-14-5-2-3-17-5#20-3-37-5#20-3-29-5#20-3-29-5

'11/9/12 (Mon)

  • Colony PCR
    • #20-#3-14-5-2-3-17-5#20-3-14-5-#2-3-17-5#1-#3-14-5-2-3-17-5#36-3-14-5-2-3-17-5-#20-3-37-5#36-3-14-5-2-3-17-5-#20-3-37-5-#20-3-37-5#2-3-11-5 Note: #36-3-14-5-2-3-17-5-#20-3-37-5-#20-3-37-5#36-3-14-5-2-3-17-5#20-3-37-5#20#29#2-3-17-5#2-3-37-5#36-3-14-5-2-3-17-5#20-3-37-5#20-3-37-5#2-9#20-3-29-5#20#36-3-14-5#2-3-17-5#36-3-14-5-2-3-17-5#20-3-37-5#20-3-37-5#20-3-14-5-2-3-17-5#1-3-14-5-2-3-17-5#36-3-14-5-2-3-17-5-20-3-37-5-#36-3-14-5-2-3-17-5#20-3-37-5#36-3-14-5-2-3-17-5#20-3-37-5#3-27-5#20#36-3-29-5#20-3-37-5

'11/9/13 (Tue)

  • Colony PCR
    • #36-3-14-5-2-3-17-5#20-3-37-5#36-3-14-5-2-3-17-5#20-3-37-5#3-27-5#20#36-3-29-5#20-3-37-5#36-3-14-5-2-3-17-5-20-3-37-5#20-3-14-5-2-3-17-5#1-3-14-5-2-3-17-5 Note: Seemes OD600 should NOT exceed 0.4~0.5, or intracellular luciferase will be saturated. *Miniprep. **#20-3-14-5-2-3-17-5#1-3-14-5-2-3-17-5#20-3-37-5#20-3-37-5#20-3-29-5#2-9#20-3-29-5#1-3-14-5-2-3-17-5#20-3-14-5-2-3-17-5#36-3-14-5-2-3-17-5-20-3-37-5#2-3-11-5#20-3-37-5#20-3-29-5#3-27-5#31#20#3-29-5#20#3-27-5#3-14-5-2-3-17-5#1-3-14-5-2-3-17-5#20-3-14-5-2-3-17-5#20-3-37-5

'11/09/14(Wed)

  • Colony PCR
    • #3-27-5#20#20-3-14-5-2-3-17-5#1-3-14-5-2-3-17-5#3-14-5#3-17-5#3-14-5-2-3-17-5#1-3-14-5-2-3-17-5#20-3-14-5-2-3-17-5#20#20#20-3-14-5-2-3-17-5#3-14-5-2-3-17-5#3-17-5#3-11-5#19#20#3-29-5#36-3-14-5-2-3-17-5#20-3-37-5#1-3-14-5-2-3-17-5#20-3-14-5-2-3-17-5#20-3-37-5#20-3-29-5#2-3-11-5#3-27-5#31

'11/09/15(Thu)

  • Colony PCR(#19#36-3-14-5-2-3-17-5-20-3-37-5#20-3-29-5#20#20-3-14-5-2-3-17-5#3-14-5-2-3-17-5#3-17-5#3-14-5#3-17-5#3-14-5-2-3-17-5#1-3-14-5-2-3-17-5#20-3-14-5-2-3-17-5#20#3-11-5#20-3-29-5#19#3-14-5-2-3-17-5#3-14-5-2-3-17-5#36-3-14-5-2-3-17-5#20-3-37-5#3-27-5#20#3-17-5#3-14-5#3-14-5-2-3-17-5#20-3-14-5-2-3-17-5#20-3-14-5-2-3-17-5

'11/09/16(Fri)

  • Colony PCR
    • sulAp(ES)-#3-14-5-2-3-17-5#3-14-5-2-3-17-5#36-3-14-5-2-3-17-5#20-3-37-5#3-27-5#20#3-17-5#3-14-5#3-14-5-2-3-17-5#20-3-14-5-2-3-17-5#36-3-14-5-2-3-17-5#20-3-37-5#3-11-5#20-3-29-5#19#3-27-5#36-3-14-5-2-3-17-5#20-3-29-5#3-27-5#36-3-14-5-2-3-17-5#20-3-14-5-2-3-17-5

'11/09/17(Sat)

  • Colony PCR (retry)
    • #3-14-5#3-17-5#36-3-14-5-2-3-17-5-20-3-37-5#3-14-5#3-17-5#20-#3-27-5#20-3-29-5

'11/09/18(Sun)

  • Colony PCR & Culture for Miniprep(#36-3-14-5-2-3-17-5-20-3-37-5#33-3-14-5-2-3-17-5#3-14-5#3-17-5#20-#3-27-5#3-14-5-2-3-17-5#31-

'11/09/19(Mon)

  • Culture for Dual luciferase assay(#36-3-14-5-2-3-17-5-20-3-37-5#33-3-14-5-2-3-17-5#1-3-14-5-2-3-17-5#20-3-14-5-2-3-17-5#36-3-14-5-2-3-17-5-20-3-37-5#33-3-14-5-2-3-17-5#24-#3-14-5-2-3-17-5#20-#3-27-5#36-3-14-5-2-3-17-5-20-3-37-5#33-3-14-5-2-3-17-5#20-3-29-5#1#20#36#33

'11/09/20(Tue)

  • colony PCR(20-3-27-5, #24-3-14-5-2-3-17-5#3-27-5 Digest(#29#20-3-29-5#3-29-5#20#3-11-5#3-14-5#3-17-5#31#36-3-29-5#20-3-37-5

'11/09/21(Wed)

  • Making plates
  • Gel extraction(#20#29#3-29-5#20-3-29-5#20#3-27-5#24#3-14-5-2-3-17-5#29#3-29-5#20-3-29-5#3-14-5-2-3-17-5#36#33-3-14-5-2-3-17-5#3-14-5-2-3-17-5#3-14-5-2-3-17-5#3-29-5#36-3-14-5-2-3-17-5

'11/09/22(Thu)

  • Ligation (retry)
    • #3-27-5#20#3-14-5-2-3-17-5-#24#29#3-29-5#20-3-29-5#3-14-5-2-3-17-5#36culture for mini prep), #33-3-14-5-2-3-17-5Maybe the amplification was failed)) *Gel extraction(#3-14-5-2-3-17-5#3-29-5#3-14-5-2-3-17-5#20-3-37-5#3-14-5-2-3-17-5#20-3-37-5#3-29-5#20-3-37-5

'11/09/23(Fri)

  • colony PCR
  • Column purification
  • Ligation
  • PCR amplification
  • Competent cell(BL21(DE3))
  • Asp diffusion test
  • Cell diffusion assay
  • Asp chemotaxis assay

'11/09/26(Mon)

  • Ligation(#20-#3-27-5#24-#3-14-5-2-3-17-5#3-29-5-#20-3-29-5-#36#3-29-5#3-29-5-#20-3-29-5-


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