Team:Tsinghua/experiments

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<h1>Protocols</h1>
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Here are our Experimental Protocols in PDF version. You're advised to view in Acrobat or Adobe Reader which you can download free from the Home Website of Adobe® Company, for other viewers we can not guarantee the right format of viewing. Those files are encrypted to keep copyrights, if you want to print them out of copy to your documents, to share with others our ideas and methods, you are welcome to ask us for the unencrypted files definitely for free~
 
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Here are our Experimental Protocols in PDF version. You're advised to view in Acrobat or Adobe Reader which you can download free from the Home Website of Adobe® Company, for other viewers we can not guarantee the right format of viewing. Those files are encrypted to keep copyrights, if you want to print them out of copy to your documents, to share with others our ideas and methods, you are welcome to ask us for the unencrypted files definitely for free~
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<font size="4"><B>Molecular Cloning</B></font>
<font size="4"><B>Molecular Cloning</B></font>
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Revision as of 15:25, 26 October 2010

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Experiment Records

July
MTWTFSS
      [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/1_July_2010&action=edit 1] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/2_July_2010&action=edit 2] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/3_July_2010&action=edit 3] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/4_July_2010&action=edit 4]
[http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/5_July_2010&action=edit 5] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/6_July_2010&action=edit 6] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/7_July_2010&action=edit 7] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/8_July_2010&action=edit 8] [http://2010.igem.org/Team:Tsinghua/Notebook/9_July_2010 9] [http://2010.igem.org/Team:Tsinghua/Notebook/10_July_2010 10] [http://2010.igem.org/Team:Tsinghua/Notebook/11_July_2010 11]
[http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/12_July_2010&action=edit 12] [http://2010.igem.org/Team:Tsinghua/Notebook/13_July_2010 13] [http://2010.igem.org/Team:Tsinghua/Notebook/14_July_2010 14] [http://2010.igem.org/Team:Tsinghua/Notebook/15_July_2010 15] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/16_July_2010&action=edit 16] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/17_July_2010&action=edit 17] [http://2010.igem.org/Team:Tsinghua/Notebook/18_July_2010 18]
[http://2010.igem.org/Team:Tsinghua/Notebook/19_July_2010 19] [http://2010.igem.org/Team:Tsinghua/Notebook/20_July_2010 20] [http://2010.igem.org/Team:Tsinghua/Notebook/21_July_2010 21] [http://2010.igem.org/Team:Tsinghua/Notebook/22_July_2010 22] [http://2010.igem.org/Team:Tsinghua/Notebook/23_July_2010 23] [http://2010.igem.org/Team:Tsinghua/Notebook/24_July_2010 24] [http://2010.igem.org/Team:Tsinghua/Notebook/25_July_2010 25]
[http://2010.igem.org/Team:Tsinghua/Notebook/26_July_2010 26] [http://2010.igem.org/Team:Tsinghua/Notebook/27_July_2010 27] [http://2010.igem.org/Team:Tsinghua/Notebook/28_July_2010 28] [http://2010.igem.org/Team:Tsinghua/Notebook/29_July_2010 29] [http://2010.igem.org/Team:Tsinghua/Notebook/30_July_2010 30] [http://2010.igem.org/Team:Tsinghua/Notebook/31_July_2010 31]
August
MTWTFSS
            [http://2010.igem.org/Team:Tsinghua/Notebook/1_August_2010 1]
[http://2010.igem.org/Team:Tsinghua/Notebook/2_August_2010 2] [http://2010.igem.org/Team:Tsinghua/Notebook/3_August_2010 3] [http://2010.igem.org/Team:Tsinghua/Notebook/4_August_2010 4] [http://2010.igem.org/Team:Tsinghua/Notebook/5_August_2010 5] [http://2010.igem.org/Team:Tsinghua/Notebook/6_August_2010 6] [http://2010.igem.org/Team:Tsinghua/Notebook/7_August_2010 7] [http://2010.igem.org/Team:Tsinghua/Notebook/8_August_2010 8]
[http://2010.igem.org/Team:Tsinghua/Notebook/9_August_2010 9] [http://2010.igem.org/Team:Tsinghua/Notebook/10_August_2010 10] [http://2010.igem.org/Team:Tsinghua/Notebook/11_August_2010 11] [http://2010.igem.org/Team:Tsinghua/Notebook/12_August_2010 12] [http://2010.igem.org/Team:Tsinghua/Notebook/13_August_2010 13] [http://2010.igem.org/Team:Tsinghua/Notebook/14_August_2010 14] [http://2010.igem.org/Team:Tsinghua/Notebook/15_August_2010 15]
[http://2010.igem.org/Team:Tsinghua/Notebook/16_August_2010 16] [http://2010.igem.org/Team:Tsinghua/Notebook/17_August_2010 17] [http://2010.igem.org/Team:Tsinghua/Notebook/18_August_2010 18] [http://2010.igem.org/Team:Tsinghua/Notebook/19_August_2010 19] [http://2010.igem.org/Team:Tsinghua/Notebook/20_August_2010 20] [http://2010.igem.org/Team:Tsinghua/Notebook/21_August_2010 21] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/22_August_2010&action=edit 22]
[http://2010.igem.org/Team:Tsinghua/Notebook/23_August_2010 23] [http://2010.igem.org/Team:Tsinghua/Notebook/24_August_2010 24] [http://2010.igem.org/Team:Tsinghua/Notebook/25_August_2010 25] [http://2010.igem.org/Team:Tsinghua/Notebook/26_August_2010 26] [http://2010.igem.org/Team:Tsinghua/Notebook/27_August_2010 27] [http://2010.igem.org/Team:Tsinghua/Notebook/28_August_2010 28] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/29_August_2010&action=edit 29]
[http://2010.igem.org/Team:Tsinghua/Notebook/30_August_2010 30] [http://2010.igem.org/Team:Tsinghua/Notebook/31_August_2010 31]
September
MTWTFSS
    [http://2010.igem.org/Team:Tsinghua/Notebook/1_September_2010 1] [http://2010.igem.org/Team:Tsinghua/Notebook/2_September_2010 2] [http://2010.igem.org/Team:Tsinghua/Notebook/3_September_2010 3] [http://2010.igem.org/Team:Tsinghua/Notebook/4_September_2010 4] [http://2010.igem.org/Team:Tsinghua/Notebook/5_September_2010 5]
[http://2010.igem.org/Team:Tsinghua/Notebook/6_September_2010 6] [http://2010.igem.org/Team:Tsinghua/Notebook/7_September_2010 7] [http://2010.igem.org/Team:Tsinghua/Notebook/8_September_2010 8] [http://2010.igem.org/Team:Tsinghua/Notebook/9_September_2010 9] [http://2010.igem.org/Team:Tsinghua/Notebook/10_September_2010 10] [http://2010.igem.org/Team:Tsinghua/Notebook/11_September_2010 11] [http://2010.igem.org/Team:Tsinghua/Notebook/12_September_2010 12]
[http://2010.igem.org/Team:Tsinghua/Notebook/13_September_2010 13] [http://2010.igem.org/Team:Tsinghua/Notebook/14_September_2010 14] [http://2010.igem.org/Team:Tsinghua/Notebook/15_September_2010 15] [http://2010.igem.org/Team:Tsinghua/Notebook/16_September_2010 16] [http://2010.igem.org/Team:Tsinghua/Notebook/17_September_2010 17] [http://2010.igem.org/Team:Tsinghua/Notebook/18_September_2010 18] [http://2010.igem.org/Team:Tsinghua/Notebook/19_September_2010 19]
[http://2010.igem.org/Team:Tsinghua/Notebook/20_September_2010 20] [http://2010.igem.org/Team:Tsinghua/Notebook/21_September_2010 21] [http://2010.igem.org/Team:Tsinghua/Notebook/22_September_2010 22] [http://2010.igem.org/Team:Tsinghua/Notebook/23_September_2010 23] [http://2010.igem.org/Team:Tsinghua/Notebook/24_September_2010 24] [http://2010.igem.org/Team:Tsinghua/Notebook/25_September_2010 25] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/26_September_2010&action=edit 26]
[http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/27_September_2010&action=edit 27] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/28_September_2010&action=edit 28] [http://2010.igem.org/Team:Tsinghua/Notebook/29_September_2010 29] [http://2010.igem.org/Team:Tsinghua/Notebook/30_September_2010 30]
October
MTWTFSS
        [http://2010.igem.org/Team:Tsinghua/Notebook/1_October_2010 1] [http://2010.igem.org/Team:Tsinghua/Notebook/2_October_2010 2] [http://2010.igem.org/Team:Tsinghua/Notebook/3_October_2010 3]
[http://2010.igem.org/Team:Tsinghua/Notebook/4_October_2010 4] [http://2010.igem.org/Team:Tsinghua/Notebook/5_October_2010 5] [http://2010.igem.org/Team:Tsinghua/Notebook/6_October_2010 6] [http://2010.igem.org/Team:Tsinghua/Notebook/7_October_2010 7] [http://2010.igem.org/Team:Tsinghua/Notebook/8_October_2010 8] [http://2010.igem.org/Team:Tsinghua/Notebook/9_October_2010 9] [http://2010.igem.org/Team:Tsinghua/Notebook/10_October_2010 10]
[http://2010.igem.org/Team:Tsinghua/Notebook/11_October_2010 11] [http://2010.igem.org/Team:Tsinghua/Notebook/12_October_2010 12] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/13_October_2010&action=edit 13] [http://2010.igem.org/Team:Tsinghua/Notebook/14_October_2010 14] [http://2010.igem.org/Team:Tsinghua/Notebook/15_October_2010 15] [http://2010.igem.org/Team:Tsinghua/Notebook/16_October_2010 16] [http://2010.igem.org/Team:Tsinghua/Notebook/17_October_2010 17]
[http://2010.igem.org/Team:Tsinghua/Notebook/18_October_2010 18] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/19_October_2010&action=edit 19] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/20_October_2010&action=edit 20] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/21_October_2010&action=edit 21] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/22_October_2010&action=edit 22] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/23_October_2010&action=edit 23] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/24_October_2010&action=edit 24]
[http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/25_October_2010&action=edit 25] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/26_October_2010&action=edit 26] [http://2010.igem.org/Team:Tsinghua/Notebook/27_October_2010 27] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/28_October_2010&action=edit 28] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/29_October_2010&action=edit 29] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/30_October_2010&action=edit 30] [http://2010.igem.org/wiki/index.php?title=Team:Tsinghua/Notebook/31_October_2010&action=edit 31]

Our Experiments were carried out by the following nine groups which are somewhat independent from each other. Here come the Groups and Their Tasks.
Group 1A：Landing pad (LP) construction and Insertion Project with Att Recomination method and Trandition Landing pad








Protocols

Here are our Experimental Protocols in PDF version. You're advised to view in Acrobat or Adobe Reader which you can download free from the Home Website of Adobe® Company, for other viewers we can not guarantee the right format of viewing. Those files are encrypted to keep copyrights, if you want to print them out of copy to your documents, to share with others our ideas and methods, you are welcome to ask us for the unencrypted files definitely for free~

Molecular Cloning
THUProtocol_1-1_Isolatio_of_plasmid_DNA

THUProtocol_1-2_DNA_Gel_Extraction

THUProtocol_1-3_Restriction_Enzyme_Digestion

THUProtocol_1-4_Ligation_of_DNA_Fragments

THUProtocol_1-5_PCR

THUProtocol_1-6_Preparation_of_Competent_Cell_for_Electro_Transformation

THUProtocol_1-7_Chemical_Transformation_of_Recombinant_DNA

THUProtocol_1-8_Electro_Transformation_of_Recombinant_DNA

Protein Isolation and Identification
THUProtocol_2-1_Protein_Isolation_for_Prokaryotes

THUProtocol_2-2_Protein_Identification_SDS-PAGE

THUProtocol_1-5_PCR

Results

Group 2(b)
1. PCR: Amplication eGFP, mCherry, Kanamycin resistant gene and Chlr four genes separatly from PIB-eGFP, PBS34, PKD13 and PKD3 plasmids.

Fan1.JPG Fan2.JPG


2. pUC19 double digestion for eGFP and Chlr, ligate with digested eGFP and Chlr separately. PCR to detect pUC19+eGFP and pUC19+ChlrPCR. All 12 samples are positive.

THU-Fan3.JPG


3. Double digest PE1 and mCherry with SalI and BamHI, double digest PC1 and Kan with KpnI and BamHI, ligate and PCR to detect. No positive results exist among PC1+K.

THU-Fan4.JPG


4. Double digest PEM2 and Kan with KpnI and BamHI as well as PC1. Ligate and using PCR to detect with universal primers. PEM2+K 1 and 5 are positive.

THU-Fan5.JPG


5. Double digest PEMK1 and Chlr with KpnI and EcoRI. Ligate and using PCR to detect with universal primers. PEMK1+Chlr 3-5 are positive.

THU-Fan6.JPG

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