User contributions for Chen Chongyi
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21 November 2007
- 06:4406:44, 21 November 2007 diff hist +21 N IGEM:Peking/2007/Chongyi Chen New page: chenchongyi@gmail.com current
26 October 2007
- 18:1618:16, 26 October 2007 diff hist +152 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. current
- 18:1418:14, 26 October 2007 diff hist +134 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!!! current
- 18:1318:13, 26 October 2007 diff hist +73 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 current
- 18:1218:12, 26 October 2007 diff hist +120 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 current
- 18:1018:10, 26 October 2007 diff hist +164 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 current
- 18:0818:08, 26 October 2007 diff hist +118 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 current
- 18:0718:07, 26 October 2007 diff hist +35 N IGEM:Peking/2007/Switch-Notebook/2007-9-20 New page: ==Pick colonies to cultivate them== current
- 18:0618:06, 26 October 2007 diff hist +80 N IGEM:Peking/2007/Switch-Notebook/2007-9-19 New page: ==Rescue of all the vectors and fragments== The result is invisible but normal. current
- 18:0618:06, 26 October 2007 diff hist +116 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== current
- 18:0518:05, 26 October 2007 diff hist +116 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== current
- 18:0418:04, 26 October 2007 diff hist 0 IGEM:Peking/2007/Switch-Notebook/2007-9-15 →Plates problme solved current
- 18:0418:04, 26 October 2007 diff hist +63 IGEM:Peking/2007/Switch-Notebook/2007-9-15 →Ligation
- 18:0318:03, 26 October 2007 diff hist +80 N IGEM:Peking/2007/Switch-Notebook/2007-9-15 New page: ==Ligation== pSB1A3 with EGFP, 434OR21, 434OR321 pSB3K3 with all the fragments
- 18:0218:02, 26 October 2007 diff hist +185 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. current
- 18:0118:01, 26 October 2007 diff hist +201 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... current
- 17:5917:59, 26 October 2007 diff hist +362 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... current
- 17:5617:56, 26 October 2007 diff hist +192 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... current
- 17:5417:54, 26 October 2007 diff hist +253 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... current
- 17:5117:51, 26 October 2007 diff hist +154 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== current
- 17:5017:50, 26 October 2007 diff hist +272 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... current
- 17:4717:47, 26 October 2007 diff hist +229 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... current
- 17:4517:45, 26 October 2007 diff hist +492 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... current
- 17:4217:42, 26 October 2007 diff hist +196 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... current
- 17:4017:40, 26 October 2007 diff hist +370 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== ... current
- 17:3617:36, 26 October 2007 diff hist +645 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 =... current
- 17:3217:32, 26 October 2007 diff hist +261 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... current
- 17:2917:29, 26 October 2007 diff hist +394 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... current
- 17:2517:25, 26 October 2007 diff hist +753 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 ... current
- 17:1817:18, 26 October 2007 diff hist +532 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... current
- 09:4309:43, 26 October 2007 diff hist +1 IGEM:Peking/2007/Switch-Notebook/2007-8-28 →transformation current
- 09:4309:43, 26 October 2007 diff hist +493 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...
- 09:3809:38, 26 October 2007 diff hist +1,026 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... current
- 09:2909:29, 26 October 2007 diff hist +16 IGEM:Peking/2007/Switch-Notebook/2007-8-12 →Miniprep and double digestion test current
- 09:2909:29, 26 October 2007 diff hist +1,340 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 ...
- 09:1709:17, 26 October 2007 diff hist +515 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... current
- 09:1109:11, 26 October 2007 diff hist +846 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... current
- 08:3608:36, 26 October 2007 diff hist +700 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... current
- 08:2408:24, 26 October 2007 diff hist +840 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 ... current
- 08:1708:17, 26 October 2007 diff hist +749 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... current
- 08:0808:08, 26 October 2007 diff hist +320 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 ... current
- 08:0408:04, 26 October 2007 diff hist +555 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 ... current
- 07:4007:40, 26 October 2007 diff hist +672 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... current
- 07:3407:34, 26 October 2007 diff hist +453 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 ... current
- 07:2807:28, 26 October 2007 diff hist +3 IGEM:Peking/2007/Switch-Notebook/2007-7-27 →Re-digestion of plx007, plx008 and plx009 current
- 07:2407:24, 26 October 2007 diff hist +960 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...
- 07:1307:13, 26 October 2007 diff hist +218 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... current
- 07:1007:10, 26 October 2007 diff hist +686 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'... current
- 07:0207:02, 26 October 2007 diff hist +664 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... current
- 06:2906:29, 26 October 2007 diff hist +138 IGEM:Peking/2007/Switch-Notebook/2007-7-18 →Colony PCR current