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• 09:44, 21 November 2007 (hist) (diff) N IGEM:Peking/2007/Chongyi Chen‎ (New page: chenchongyi@gmail.com) (top)
• 21:16, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-26‎ (New page: ==Miniprep== EGFP-I7100 in two tubes OR21-I7100 and OR321-I7100 one tube each This is the base for the next liagtion step, just like previous vectors.) (top)
• 21:14, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-25‎ (New page: ==double digestion== EGFP plsmid wrong, all the other ligation plasmid seems to be right! Thus our first step in ligation is done!!!) (top)
• 21:13, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-24‎ (New page: ==Rescue from gel== cI 434 and cI is visible. OR21, OR321 and EGFP fail) (top)
• 21:12, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-23‎ (New page: ==Double digestion== all the vectors ==Positive transformation== The first ligation product: I7100 with OR21/321/EGFP) (top)
• 21:10, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-22‎ (New page: ==PCR of cI and cI434== I got the fragments. system: template 1uL Ex buffer 5uL dNTP 5uL primer 1uL each Ex Taq 0.25uL Add water to make a total 50uL system) (top)
• 21:08, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-21‎ (New page: ==Double digestion test== plasmid 4uL EcoRI/PstI 0.5uL each Buffer H 2uL ddH2O 13uL A total 20uL digestion system) (top)
• 21:07, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-20‎ (New page: ==Pick colonies to cultivate them==) (top)
• 21:06, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-19‎ (New page: ==Rescue of all the vectors and fragments== The result is invisible but normal.) (top)
• 21:06, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-18‎ (New page: ==Miniprep of pSB1A3, PSB3K3== ==double digestion of the two vectors== ==double digestion of all other fragments==) (top)
• 21:05, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-17‎ (New page: ==Miniprep of pSB1A3, PSB3K3== ==double digestion of the two vectors== ==double digestion of all other fragments==) (top)
• 21:04, 26 October 2007 (hist) (diff) IGEM:Peking/2007/Switch-Notebook/2007-9-15‎ (→Plates problme solved) (top)
• 21:04, 26 October 2007 (hist) (diff) IGEM:Peking/2007/Switch-Notebook/2007-9-15‎ (→Ligation)
• 21:03, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-15‎ (New page: ==Ligation== pSB1A3 with EGFP, 434OR21, 434OR321 pSB3K3 with all the fragments)
• 21:02, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-14‎ (New page: ==miniprep of pSB3K3 and pSB1A3== re-digestion of these two vectors overnight ==Struggling with the problem of plates== I think the antibiotics such as Kan and Amp may be the problem.) (top)
• 21:01, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-13‎ (New page: ==Cultivation== I7100 and I13521, two main vectors in our plan ==Trying to solve the problem of wrong plates== ==Colony PCR== Fail! Because the plates are wrong so the colonies are wron...) (top)
• 20:59, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-12‎ (New page: ==Problem and Warning== Our Kana and Amp plates seems all wrong!!! I have discarded them! ==Ligation== pSB3K3 with so many fragments ==Transformation== The previous ligation system. Al...) (top)
• 20:56, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-11‎ (New page: ==Positive transformation of pSB1A3== ==RS1-EGFP precipitation and rescued== ==Tansformation of the ligationi system last night== ==PCR 434OR321/21/EGFP== This time use ex-taq and 35 cy...) (top)
• 20:54, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-10‎ (New page: ==Colony PCR== Finally we have some colonies that may turn out right ==Rescue by precipitation of all fragments== ==Ex-Taq PCR of RS1 and EGFP== We got the two fragments ==ligation== I7...) (top)
• 20:51, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-9‎ (New page: ==Another ligation== The only thing we can do is to ligate I7100 with 434OR321/21 and EGFP. ==Fragments digestion and precipitation== ==Transformation==) (top)
• 20:50, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-8‎ (New page: ==Ligation of I7100 with 434OR321, 434OR21 and EGFP== Result: no colonies visible overnight Analysis: The plate may still have a problem! ==Colony PCR I7100 with OR321== That is the only...) (top)
• 20:47, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-7‎ (New page: ==Rescue by precipitation of cIind-== ==double digestion of the fragments== ==rescue by gel of I7100== ==Ligation and transformation for a second time== ==Temperature Grad PCR== To ens...) (top)
• 20:45, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-6‎ (New page: ==Check our plate== By smearing competent cells directly onto plates without any transformation procedure, we can check whether the plates with Kanamycin is right. Result: The plates have...) (top)
• 20:42, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-5‎ (New page: ==Colony PCR== To check I7100 ligation with EGFP, 434OR21, 434OR321 ==Rescue from gel of I7100 vector after digestion== But the rescue result is not good. The band seems weird! We will d...) (top)
• 20:40, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-4‎ (New page: ==Rescue I7100 by gel== ==Ligation of I7100 with 434OR21/321/EGFP== ==digestion test of T1TE-T1T2-pcc010== This time I use XhoI and KpnI in M buffer ==COlony PCR of I7100-434OR, EGFP== ...) (top)
• 20:36, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-3‎ (New page: ==Precipitation of EGFP, 434OR21/321== rescue them afterwards and run a gel to check ==PCR double repressors== This is a second time, just in case we used up all the products last time =...) (top)
• 20:32, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-9-2‎ (New page: ==PCR fragments for next step in constructio== SS1, SS2, SS3, SD1, SD2, SD3, rec, RD1, CD1, RS1, GFP ==add a control== self-ligation of I7100 ==digestion== EGFP, cI434OR21/321 using Eco...) (top)
• 20:29, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-8-31‎ (New page: ==Colony PCR of T1TE-T1T2-pcc010== primer: pcctestR and pkan-Kan-F some colonies may be right! ==Colony PCR of 434OR/21/321== primer: MBF and MBR some colonies may be right, send to seq...) (top)
• 20:25, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-8-30‎ (New page: ==Cultivate ''E.coli''== bacteria with sulA, RS2, RD3-plx012 bacteria with pSB3K3, pSB1A3 and pcc056 ==PCR 434OAR321 and 434OR21== PCR system: Ex-Taq buffer 5uL dNTP 5uL template 1uL ...) (top)
• 20:18, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-8-29‎ (New page: ==Preparation of liquid medium LB and LB Amp== ==miniprep and digestion test== sulA, RS2, RD3-plx012 plasmid is isolated. Double digestion system: H buffer 2uL plasmid 7uL XhoI/SalI 0...) (top)
• 12:43, 26 October 2007 (hist) (diff) IGEM:Peking/2007/Switch-Notebook/2007-8-28‎ (→transformation) (top)
• 12:43, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-8-28‎ (New page: ==Colony PCR of CD2, SulA, RS2, RD3 ligated with plx012== buffer 1.5uL dNTP 1.5uL dH2O 11uL test-F/GFP-test-R 0.5uL each easy taq 0.1uL elongation time span of 1 minute ==T1T2-pcc01...)
• 12:38, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-8-13‎ (New page: ==inactivate T1TE-pcc010== using phenol/choloroform to inactivate NotI/XhoI enzyme rescue by precipitation ==T1T2-pcc010 digestion test== using SacII and T buffer with BSA in it ==re-PC...) (top)
• 12:29, 26 October 2007 (hist) (diff) IGEM:Peking/2007/Switch-Notebook/2007-8-12‎ (→Miniprep and double digestion test) (top)
• 12:29, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-8-12‎ (New page: ==rescue by precipitation of fragment after digestion== electrophoresis result to show that SD2, SD3 and Prec successfully ligated. ==Colony PCR of T1T2-pcc010== easy Taq 0.1uL Pkan-F ...)
• 12:17, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-8-10‎ (New page: ==rescue by precipitation== 9 fragments: lac, rec, sul, SS1-3, SD1-3, digested by XhoI/SpeI 1 vector: GFP-plx007, digested by XhoI/XbaI ==miniprep== T1TE-plx007 T1TE-T1T2-pcc010 double...) (top)
• 12:11, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-8-9‎ (New page: ==Colony PCR of SD-pcc002, EGFP-pcc002== ==miniprep of T1TE-plx010 and GFP-plx003== digestion test: T1TE-plx010 10uL ScaI/XhoI 0.5uL each H buffer 2uL add water to a total 20uL GFP-p...) (top)
• 11:36, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-8-8‎ (New page: ==Colony PCR== T1TE-plx010 using the primer T1TE-F/R result: not very good, the T1TE fragment seems to be smaller than expected. GFP-plx003 using GFP-F/R result: GFP colony PCR has no p...) (top)
• 11:24, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-8-2‎ (New page: ==CD2-T and T-1 send to sequencing== ==T1TE and T1T2== T1TE digestion by ClaI/SalI and T1T2 digestion by NotI/XhoI, then rescued. ==LacZa-plx009 miniprep== NdeI and XhoI digestion using ...) (top)
• 11:17, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-8-1‎ (New page: ==transformation== lacZa-plx009 gel rescue, lacZa, lacZa-pcc009 precipitation rescue, plx009 and lacZa precipitation rescue ==CD2-T sequencing result== wrong! We have to re-PCR CD2 ==RD2...) (top)
• 11:08, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-7-31‎ (New page: ==re-digestion of plx007/008/009== Only Plx009 seems to be right! ==double digestion of plx009-lacZa== plx009 4uL buffer H 6uL XhoI/SalI 2uL each H2O 46uL Total 60uL system ==Colony ...) (top)
• 11:04, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-7-30‎ (New page: ==transformation of MCS-pcc009and pcc010== Smear the transformation onto Cm+LB plate ==plx007,008,009 positive transformation== Pick colonies ==colony PCR of RD2-pGEM-T== using Primer ...) (top)
• 10:40, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-7-29‎ (New page: ==CD2-T send to sequencing== after colony PCR and double digestion test result is right ==Rescue of Pla-T1T2, Pla-T1TE by electrophoresis== test result after rescue: good with high conce...) (top)
• 10:34, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-7-28‎ (New page: ==rescue of digestion production of XhoI/SalI== ==double digestion test of plx008/009-lacZa== using two systems of ScaI/SalI and XhoI/ScaI eletrophoresis to check the result ==Pick 12 ...) (top)
• 10:28, 26 October 2007 (hist) (diff) IGEM:Peking/2007/Switch-Notebook/2007-7-27‎ (→Re-digestion of plx007, plx008 and plx009) (top)
• 10:24, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-7-27‎ (New page: ==Re-digestion of plx007, plx008 and plx009== Plasmid 2uL XhoI 0.5uL H buffer 2uL ddH2O 15.5uL electrophoresis result: invisible ==pick colony of plx008, plx009== ==rescue by precipitat...)
• 10:13, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-7-22‎ (New page: ==RS fragment rescue== electrophoresis result: thanks to temperature grad, RS1, RS2 and RS3 are all got ==transformation of pRec7== ==cultivation of pRec7 E.coli== ==LB medium preparat...) (top)
• 10:10, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-7-21‎ (New page: ==PCR RS1,2,3== ddH2O 36.75uL Buffer 5uL dNTP 5uL pRec7-8 1uL Rec-S-1-F 1uL Rec-S-1-Rnew 1uL Ex-Taq 0.25uL total 50uL temperature grad: 53.2, 58.5, 63.5, 68.1 result: RS1 and RS2'...) (top)
• 10:02, 26 October 2007 (hist) (diff) N IGEM:Peking/2007/Switch-Notebook/2007-7-20‎ (New page: ==Miniprep of pLx007-lacZa(8)== ==double digestion test of lacZa-plx007/008/009== ===method I=== H buffer 2uL XhoI/SalI 0.5uL each plasmid 12uL dH2O 5uL total 20uL Failed! ===Method...) (top)
• 09:29, 26 October 2007 (hist) (diff) IGEM:Peking/2007/Switch-Notebook/2007-7-18‎ (→Colony PCR) (top)

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