Team:KAIT Japan/Notebook

From 2014.igem.org

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__NOTOC__
__NOTOC__
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='''Notebook'''=
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=<font size ="7">Notebook</font>=
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:We made an experiment every day. It was process at trial and error.
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<br>
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=='''Basic task'''==
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<br>
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:<big>'''Date:8/18'''</big> Miniprep
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-
=='''Creating parts of HlyA and GFP'''==
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=='''<font size="6">Creating parts of GFP and HlyA</font>'''==
:<big>'''Date:8/22'''</big> The refinement of DNA and PCR and Electrophoresis
:<big>'''Date:8/22'''</big> The refinement of DNA and PCR and Electrophoresis
:<big>'''Date:8/25'''</big> PCR and Electrophoresis and Restriction
:<big>'''Date:8/25'''</big> PCR and Electrophoresis and Restriction
:<big>'''Date:8/26'''</big> Insert gene into TAvectar and Transformation
:<big>'''Date:8/26'''</big> Insert gene into TAvectar and Transformation
:<big>'''Date:8/26'''</big> Blue white Selection and Electrophoresis(8/25 Restriction) and DNA Extraction
:<big>'''Date:8/26'''</big> Blue white Selection and Electrophoresis(8/25 Restriction) and DNA Extraction
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=='''Creating parts of IL-10α,IL-10β,STAT3'''==
+
:<big>'''Date:8/27'''</big> Electrophoresed the DNA that We did refinement on August 26 to confirmed it
 +
:<big>'''Date:8/28'''</big> Check the HlyA and GFP for colony PCR
 +
:<big>'''Date:8/29'''</big> DNA purify the HlyA and GFP
 +
:<big>'''Date:8/29'''</big> Ligation product and check for PCR
 +
:<big>'''Date:9/1'''</big> Build replica the HlyA and GFP
 +
:<big>'''Date:9/1'''</big> Restriction enzyme treatment the HlyA and GFP
 +
:<big>'''Date:9/2'''</big> Restriction enzyme treatment the HlyA and GFP in 9/1 was NANOSEP
 +
:<big>'''Date:9/3'''</big> Ligation and PCR, DNA purification the HlyA and GFP
 +
:<big>'''Date:9/4'''</big> Restriction enzyme treatment the HlyA and GFP
 +
:<big>'''Date:9/5'''</big> Restriction enzyme treatment the vector
 +
:<big>'''Date:9/5'''</big> Ligation the vector and insert. So transformation.
 +
:<big>'''Date:9/9'''</big> Check the HiyA and GFP replica.Ligation and transformation the vector and insert DNA.
 +
:<big>'''Date:9/10'''</big> Transformation in 9/9 check the colony PCR.TA cloning the HlyA+GFP.
 +
:<big>'''Date:9/11'''</big> Ligation the vector and insert DNA.
 +
:<big>'''Date:9/12~9/15'''</big> Transformation the Ligation product.
 +
:<big>'''Date:9/16'''</big> Noticed wrong the primer.so restart from the First.
 +
:<big>'''Date:9/17'''</big> PCR the DNA abstracted for iGEM kit.It was TA cloning and restriction enzyme treatment.
 +
:<big>'''Date:9/18'''</big> Ligation the GFP and HlyA.
 +
:<big>'''Date:9/19'''</big> Electrophoresis the ligation the GFP+HlyA in 9/18
 +
:<big>'''Date:9/20'''</big> DNA purify and PCR
 +
:<big>'''Date:9/21'''</big> PCR and DNA purify
 +
:<big>'''Date:9/22'''</big> Ligation the GFP+HlyA
 +
:<big>'''Date:9/23'''</big> PCR and DNA purify
 +
:<big>'''Date:9/24'''</big> TA Cloning was GFP and HlyA
 +
:<big>'''Date:9/26'''</big> Restriction enzyme treatment the ligation the GFP+HlyA in 9/22
 +
:<big>'''Date:9/27'''</big> Check the TA cloning of GFP+HlyA
 +
:<big>'''Date:9/29'''</big> Restriction enzyme treatment the GFP+HlyA and the pSBIC3
 +
:<big>'''Date:9/29'''</big> The vector and the insert were NANOSEP and lectrophoresis
 +
:<big>'''Date:9/30'''</big> The vector and the insert were ligation and transformation
 +
:<big>'''Date:9/30'''</big> The GFP+HlyA was PCR and DNA purify
 +
:<big>'''Date:10/1'''</big> Electrophoresis the GFP+HlyA was PCR and DNA purify in 9/30
 +
:<big>'''Date:10/2'''</big> Restriction enzyme treatment and ligation the GFP+HlyA
 +
:<big>'''Date:10/3'''</big> Check the ligation in 10/2
 +
:<big>'''Date:10/3'''</big> Transformation the plasmid
 +
:<big>'''Date:10/5'''</big> Colony PCR
 +
:<big>'''Date:10/6'''</big> Miniprep and Electrophoresis
 +
 
 +
 
 +
<br>
 +
<br>
 +
 
 +
=='''<font size="6">Creating parts of STAT3</font>'''==
:<big>'''Date:8/19'''</big> PCR(8/18 Miniprep) and Electrophoresis
:<big>'''Date:8/19'''</big> PCR(8/18 Miniprep) and Electrophoresis
-
:<big>'''Date:8/20'''</big> PCR(Lowered 2℃ from Tm value.) and Electrophoresis
+
:<big>'''Date:8/20~8/22'''</big> PCR(Lowered 2℃ from Tm value.) and Electrophoresis
-
:<big>'''Date:8/21'''</big> PCR and Electrophoresis
+
-
:<big>'''Date:8/22'''</big> PCR and Electrophoresis
+
:<big>'''Date:8/25'''</big> PCR and Electrophoresis(Changed to Taq HOTSTER from EX taq.)
:<big>'''Date:8/25'''</big> PCR and Electrophoresis(Changed to Taq HOTSTER from EX taq.)
:<big>'''Date:8/26'''</big> PCR and Electrophoresis(Using a mutation primer.)
:<big>'''Date:8/26'''</big> PCR and Electrophoresis(Using a mutation primer.)
 +
:<big>'''Date:8/27'''</big> First Variation introduction (with Prime STAR Max)
 +
:<big>'''Date:9/2'''</big> PCR and Electrophoresis
 +
:<big>'''Date:9/5~9/8'''</big> PCR and Electrophoresis
 +
:<big>'''Date:9/9'''</big> Sequence(to To confirm whether variation happened in DNA of STAT3 )
 +
:<big>'''Date:9/10~9/12'''</big> PCR and Electrophoresis
 +
:<big>'''Date:9/13~9/15'''</big> DNA purification and electrophoresis
 +
:<big>'''Date:9/16~9/17'''</big> DNA purification and colony PCR(Failure)
 +
:<big>'''Date:9/18'''</big> colony PCR(Failure)
 +
:<big>'''Date:9/19~9/22'''</big> colony PCR and electrophoresis
 +
:<big>'''Date:9/23'''</big> PCR and Electrophoresis(We found out that DNA polymerase was malfunction)
 +
:<big>'''Date:9/24'''</big> PCR and  Electrophoresis and DNA DNA purification
 +
:<big>'''Date:9/25'''</big> PCR(to find annealing temperature)
 +
:<big>'''Date:9/26~9/30'''</big> DNA purification and electrophoresis(to use DNA for the sequence)
 +
:<big>'''Date:10/1'''</big> Dilution after Concentration measurement of DNA
 +
:<big>'''Date:10/2'''</big> Sequence(to check mutation of DNA)/The variation was not found.
 +
:<big>'''Date:10/4'''</big> PCR and electrophoresis
 +
:<big>'''Date:10/5~10/7'''</big> Colony PCR and DNA purification
 +
:<big>'''Date:10/8'''</big> Concentration measurement of DNA
 +
:<big>'''Date:10/9'''</big> Sequence(to check mutation of DNA)/The variation was not found
 +
 +
 +
 +
<br>
 +
<br>
 +
 +
=='''<font size="6">Creating parts of IL-10α,IL-10β</font>'''==
 +
:<big>'''Date:8/19'''</big> PCR(8/18 Miniprep) and Electrophoresis
 +
:<big>'''Date:8/20~8/22'''</big> PCR(Lowered 2℃ from Tm value.) and Electrophoresis
 +
:<big>'''Date:8/25'''</big> PCR and Electrophoresis(Changed to Taq HOTSTER from EX taq.)
 +
:<big>'''Date:8/26'''</big> PCR and Electrophoresis(Using a mutation primer.)
 +
:<big>'''Date:8/27'''</big> First Variation introduction (with Prime STAR Max)
 +
:<big>'''Date:9/2'''</big> PCR
 +
:<big>'''Date:9/4'''</big> Transformation
 +
:<big>'''Date:9/5'''</big> Colony PCR
 +
:<big>'''Date:9/7'''</big> ColonyPCR
 +
:<big>'''Date:9/8'''</big> DNA extraction and Sequence(to To confirm whether variation happened in DNA of IL-10β)
 +
:<big>'''Date:9/9'''</big> DNA sequence
 +
:<big>'''Date:9/10'''</big> PCR and Electtrophoresis
 +
:<big>'''Date:9/11'''</big> Colony PCR
 +
:<big>'''Date:9/12'''</big> DNA extraction
 +
:<big>'''Date:9/13'''</big> PCR and Electrophoresis
 +
:<big>'''Date:9/15'''</big> ultrafiltration
 +
:<big>'''Date:9/16'''</big> DNA sequence(to comfirm whether variation happened in DNA of IL-10β)
 +
:<big>'''Date:9/20'''</big> PCR
 +
:<big>'''Date:9/23'''</big> Colony PCR
 +
:<big>'''Date:9/24'''</big> Electrophoresis
 +
:<big>'''Date:9/26'''</big> ultrafiltration
 +
:<big>'''Date:9/27'''</big> PCR and colony PCR
 +
:<big>'''Date:9/29'''</big> Electrophoresis
 +
:<big>'''Date:9/30'''</big> DNA Extraction
 +
:<big>'''Date:10/1'''</big> Colony PCR and DNA extraction
 +
:<big>'''Date:10/2'''</big> Electrophoresis
 +
:<big>'''Date:10/3'''</big> PCR(using Prime Star MAX Premix) and Electrophoresis
 +
:<big>'''Date:10/4'''</big> PCR(using Prime Star MAX Premix) and Electrophoresis
 +
:<big>'''Date:10/5'''</big> Electrophoresis
 +
 +
<br>
 +
<br>
 +
 +
=='''<font size="6">Creating parts of HRV,IL-5Ra,IL-5Rb,AraC,Arac Promoter</font>'''==
 +
:<big>'''Date:9/16'''</big> PCR:Ara,AraPro,HRV  Transformation:IL-5Ra,IL-5Rb
 +
:<big>'''Date:9/17'''</big> Restriction enzyme treatment and  insert to TA vector and transformation
 +
:<big>'''Date:9/18'''</big> transformation
 +
:<big>'''Date:9/19'''</big> miniprep and colony PCR
 +
:<big>'''Date:9/20~9/22'''</big> PCR
 +
:<big>'''Date:9/23'''</big> TA Cloning
 +
:<big>'''Date:9/24'''</big> TA Cloning and PCR
 +
:<big>'''Date:9/25'''</big> Restriction enzyme treatment and PCR
 +
:<big>'''Date:9/26'''</big> Colony PCR and Ligation
 +
:<big>'''Date:9/27'''</big> TA Cloning and Ligation and colony PCR
 +
:<big>'''Date:9/28'''</big> PCR
 +
:<big>'''Date:9/29'''</big> colony PCR and PCR and DNA refinement and Ligation
 +
:<big>'''Date:9/30'''</big> DNA refinement and Ligation
 +
:<big>'''Date:10/1'''</big> Restriction enzyme treatment and Ligation
 +
:<big>'''Date:10/2'''</big> PCR and Ligation and DNA refinement
 +
:<big>'''Date:10/3'''</big> Ligation
 +
:<big>'''Date:10/4'''</big> Ligation
 +
:<big>'''Date:10/7'''</big> PCR
 +
:<big>'''Date:10/8'''</big> PCR and Restriction enzyme
 +
:<big>'''Date:10/9'''</big> DNA refinement  and Restriction enzyme
 +
:<big>'''Date:10/10'''</big> Phoshatase treatment
 +
:<big>'''Date:10/11'''</big> DNA refinement and Ligation and Restriction enzyme treatment
 +
:<big>'''Date:10/12'''</big> DNA refinement and PCR and Ligation and phoshatase treatment
 +
 +
 +
 +
 +
 +
|}
|}

Latest revision as of 09:44, 17 October 2014

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Notebook



Creating parts of GFP and HlyA

Date:8/22 The refinement of DNA and PCR and Electrophoresis
Date:8/25 PCR and Electrophoresis and Restriction
Date:8/26 Insert gene into TAvectar and Transformation
Date:8/26 Blue white Selection and Electrophoresis(8/25 Restriction) and DNA Extraction
Date:8/27 Electrophoresed the DNA that We did refinement on August 26 to confirmed it
Date:8/28 Check the HlyA and GFP for colony PCR
Date:8/29 DNA purify the HlyA and GFP
Date:8/29 Ligation product and check for PCR
Date:9/1 Build replica the HlyA and GFP
Date:9/1 Restriction enzyme treatment the HlyA and GFP
Date:9/2 Restriction enzyme treatment the HlyA and GFP in 9/1 was NANOSEP
Date:9/3 Ligation and PCR, DNA purification the HlyA and GFP
Date:9/4 Restriction enzyme treatment the HlyA and GFP
Date:9/5 Restriction enzyme treatment the vector
Date:9/5 Ligation the vector and insert. So transformation.
Date:9/9 Check the HiyA and GFP replica.Ligation and transformation the vector and insert DNA.
Date:9/10 Transformation in 9/9 check the colony PCR.TA cloning the HlyA+GFP.
Date:9/11 Ligation the vector and insert DNA.
Date:9/12~9/15 Transformation the Ligation product.
Date:9/16 Noticed wrong the primer.so restart from the First.
Date:9/17 PCR the DNA abstracted for iGEM kit.It was TA cloning and restriction enzyme treatment.
Date:9/18 Ligation the GFP and HlyA.
Date:9/19 Electrophoresis the ligation the GFP+HlyA in 9/18
Date:9/20 DNA purify and PCR
Date:9/21 PCR and DNA purify
Date:9/22 Ligation the GFP+HlyA
Date:9/23 PCR and DNA purify
Date:9/24 TA Cloning was GFP and HlyA
Date:9/26 Restriction enzyme treatment the ligation the GFP+HlyA in 9/22
Date:9/27 Check the TA cloning of GFP+HlyA
Date:9/29 Restriction enzyme treatment the GFP+HlyA and the pSBIC3
Date:9/29 The vector and the insert were NANOSEP and lectrophoresis
Date:9/30 The vector and the insert were ligation and transformation
Date:9/30 The GFP+HlyA was PCR and DNA purify
Date:10/1 Electrophoresis the GFP+HlyA was PCR and DNA purify in 9/30
Date:10/2 Restriction enzyme treatment and ligation the GFP+HlyA
Date:10/3 Check the ligation in 10/2
Date:10/3 Transformation the plasmid
Date:10/5 Colony PCR
Date:10/6 Miniprep and Electrophoresis




Creating parts of STAT3

Date:8/19 PCR(8/18 Miniprep) and Electrophoresis
Date:8/20~8/22 PCR(Lowered 2℃ from Tm value.) and Electrophoresis
Date:8/25 PCR and Electrophoresis(Changed to Taq HOTSTER from EX taq.)
Date:8/26 PCR and Electrophoresis(Using a mutation primer.)
Date:8/27 First Variation introduction (with Prime STAR Max)
Date:9/2 PCR and Electrophoresis
Date:9/5~9/8 PCR and Electrophoresis
Date:9/9 Sequence(to To confirm whether variation happened in DNA of STAT3 )
Date:9/10~9/12 PCR and Electrophoresis
Date:9/13~9/15 DNA purification and electrophoresis
Date:9/16~9/17 DNA purification and colony PCR(Failure)
Date:9/18 colony PCR(Failure)
Date:9/19~9/22 colony PCR and electrophoresis
Date:9/23 PCR and Electrophoresis(We found out that DNA polymerase was malfunction)
Date:9/24 PCR and Electrophoresis and DNA DNA purification
Date:9/25 PCR(to find annealing temperature)
Date:9/26~9/30 DNA purification and electrophoresis(to use DNA for the sequence)
Date:10/1 Dilution after Concentration measurement of DNA
Date:10/2 Sequence(to check mutation of DNA)/The variation was not found.
Date:10/4 PCR and electrophoresis
Date:10/5~10/7 Colony PCR and DNA purification
Date:10/8 Concentration measurement of DNA
Date:10/9 Sequence(to check mutation of DNA)/The variation was not found




Creating parts of IL-10α,IL-10β

Date:8/19 PCR(8/18 Miniprep) and Electrophoresis
Date:8/20~8/22 PCR(Lowered 2℃ from Tm value.) and Electrophoresis
Date:8/25 PCR and Electrophoresis(Changed to Taq HOTSTER from EX taq.)
Date:8/26 PCR and Electrophoresis(Using a mutation primer.)
Date:8/27 First Variation introduction (with Prime STAR Max)
Date:9/2 PCR
Date:9/4 Transformation
Date:9/5 Colony PCR
Date:9/7 ColonyPCR
Date:9/8 DNA extraction and Sequence(to To confirm whether variation happened in DNA of IL-10β)
Date:9/9 DNA sequence
Date:9/10 PCR and Electtrophoresis
Date:9/11 Colony PCR
Date:9/12 DNA extraction
Date:9/13 PCR and Electrophoresis
Date:9/15 ultrafiltration
Date:9/16 DNA sequence(to comfirm whether variation happened in DNA of IL-10β)
Date:9/20 PCR
Date:9/23 Colony PCR
Date:9/24 Electrophoresis
Date:9/26 ultrafiltration
Date:9/27 PCR and colony PCR
Date:9/29 Electrophoresis
Date:9/30 DNA Extraction
Date:10/1 Colony PCR and DNA extraction
Date:10/2 Electrophoresis
Date:10/3 PCR(using Prime Star MAX Premix) and Electrophoresis
Date:10/4 PCR(using Prime Star MAX Premix) and Electrophoresis
Date:10/5 Electrophoresis



Creating parts of HRV,IL-5Ra,IL-5Rb,AraC,Arac Promoter

Date:9/16 PCR:Ara,AraPro,HRV Transformation:IL-5Ra,IL-5Rb
Date:9/17 Restriction enzyme treatment and insert to TA vector and transformation
Date:9/18 transformation
Date:9/19 miniprep and colony PCR
Date:9/20~9/22 PCR
Date:9/23 TA Cloning
Date:9/24 TA Cloning and PCR
Date:9/25 Restriction enzyme treatment and PCR
Date:9/26 Colony PCR and Ligation
Date:9/27 TA Cloning and Ligation and colony PCR
Date:9/28 PCR
Date:9/29 colony PCR and PCR and DNA refinement and Ligation
Date:9/30 DNA refinement and Ligation
Date:10/1 Restriction enzyme treatment and Ligation
Date:10/2 PCR and Ligation and DNA refinement
Date:10/3 Ligation
Date:10/4 Ligation
Date:10/7 PCR
Date:10/8 PCR and Restriction enzyme
Date:10/9 DNA refinement and Restriction enzyme
Date:10/10 Phoshatase treatment
Date:10/11 DNA refinement and Ligation and Restriction enzyme treatment
Date:10/12 DNA refinement and PCR and Ligation and phoshatase treatment