Team:SSTi-SZGD/js/date

var Json = { "March":{ "D27":"1.The production of competent cells by Kit", "D28":"1.SDS-PAGE
2.T-vector transformation", "D29":"1.GDNA was extracted by CTAB
2.T1 plasmid was inoculated
3.Stock" }, "April":{ "D5":"1.Plasmid DNA extraction
2.T2 generation plasmid inoculation", "D6":"1.Pave board
2.Inoculate to LB liquid medium", "D7":"1.extract gDNA", "D10":"1.GDNA was extracted by CTAB", "D11":"1.Extraction of yeast gDNA by CTAB", "D12":"1.Agarose gel electrophoresis", "D13":"1.Cloning experiments of Control DNA fragments", "D17":"1.Extraction of yeast gDNA by CTAB", "D18":"1.Agarose gel electrophoresis", "D19":"1.Extraction plasmid
2.Agarose gel electrophoresis
3.AMP conversion plate validation", "D20":"1.Cloning experiments of Control DNA fragments
2.Extraction of yeast gDNA by CTAB", "D21":"1.Agarose gel electrophoresis", "D24":"1.Cloning experiments of Control DNA fragments
2.Extraction plasmid
3.Agarose gel electrophoresis
4.PCR", "D25":"1.Purification of GDNA
2.Inoculated taste bacteria fluid
3.Agarose gel electrophoresis
4.PCR", "D26":"1.inoculation
2.Transformation of preparing competent cells and competent
3.Agarose gel electrophoresis", "D27":"1.Transformed plate observation
2.Enzyme digestion was performed with a kit
3.Agarose gel electrophoresis
4.PCR", "D28":"1.Extraction plasmid
2.Agarose gel electrophoresis" }, "May":{ "D1":"1.Transformed light off plasmid
2.Stock", "D2":"1.Light off transformed plates were observed
2.Stock", "D3":"1.To configure Streptomycin
2.Plasmid connection
3.Plasmid purification", "D4":"1.Extraction plasmid
2.Inoculate to LB liquid medium", "D5":"1.Streptomycin gradient plate observation
2.Lightoff plasmid was extracted
3.Agarose gel electrophoresis", "D7":"1.Streptomycin test
2.Do light on system validation", "D9":"1.Extraction plasmid
2.Agarose gel electrophoresis
3.Stock
4.inoculation
5.light on T1 generation transformation", "D10":"1.Extraction plasmid
2.Agarose gel electrophoresis
3.Rubber cutting
4.Agarose gel electrophoresis", "D11":"1.Agarose gel electrophoresis plastic recycling
2.Agarose gel electrophoresis", "D15":"1.Stock
2.Agarose gel electrophoresis plastic recycling
3.Agarose gel electrophoresis", "D16":"1.Agarose gel electrophoresis plastic recycling
2.Agarose gel electrophoresis
3.To configure Streptomycin
4.light on T1 generation transformation", "D17":"1.light on T1 generation transformation
2.Stock" }, "June":{ "D26":"1.Learning HPLC operations
2.Enzyme digestion", "D27":"1.Rubber cutting
2.Agarose gel electrophoresis", "D28":"1.Agarose gel electrophoresis plastic recycling
2.Agarose gel electrophoresis", "D29":"1.Enzyme digestion
2.Agarose gel electrophoresis", "D30":"1.Agarose gel electrophoresis plastic recycling
2.Agarose gel electrophoresis" }, "July":{ "D1":"1.Mutation plevion mCherry the first site of the plasmid DNA.
•Mutant Strand Synthesis Reaction (Thermal Cycling)
•Dpn I Digestion of the Amplification Products
•Transformation of XL10-Gold Ultracompetent Cell", "D2":"1.Transformation(7/1) of DH5a competent Cells", "D3":"1.Purified strains(7/2)
•Using LB /90X and 100X Penicillin-Streptomycin solution plate culture medium (according to the nature of the bacteria by adding appropriate Penicillin-Streptomycin Solution), with the inoculation of E. coli, in the flat plate culture on the grading, to be able to appear a single colony is appropriate.
•The above-mentioned flatbed culture medium was placed in a constant temperature incubator at 37℃ overnight,about 12h, this is T2 period.", "D4":"1.Plasmid DNA extraction(7/3)
•Using Takara extraction kit", "D5":"1.The mutated plasmid was sequenced", "D8":"1.Mutation plevion mCherry the second site of the plasmid DNA.
•Transformation(7/4) of DH5a competent Cells", "D9":"1.Purified strains(7/8)
•Using LB /90X and 100X Penicillin-Streptomycin solution plate culture medium (according to the nature of the bacteria by adding appropriate Penicillin-Streptomycin Solution), with the inoculation of E. coli, in the flat plate culture on the grading, to be able to appear a single colony is appropriate.
•The above-mentioned flatbed culture medium was placed in a constant temperature incubator at 37℃ overnight,about 12h, this is T2 period.", "D10":"1.Plasmid DNA extraction(7/9)
•Using Takara extraction kit", "D11":"1.The mutated plasmid was sequenced", "D14":"1.Mutation plevion mCherry the second site of the plasmid DNA.
•Transformation(7/10) of DH5a competent Cells", "D15":"1.Mutation plevion mCherry the first site of the plasmid DNA.
•Mutant Strand Synthesis Reaction (Thermal Cycling)
•Dpn I Digestion of the Amplification Products
•Transformation of XL10-Gold Ultracompetent Cells", "D16":"1.Plasmid DNA extraction(7/15)
•Using Takara extraction kit", "D17":"1.The mutated plasmid was sequenced", "D20":"1.Cultivation Mhei-mcherry gene
•A typical colony in the mhei-mcherry plate was streaked on streptomycin (800 ug / ml) 3 ml LB liquid medium and incubated overnight at 37 ℃.", "D21":"1.Cultivation Mhei-mcherry gene protein
•Measuring OD600 = 1.146, batch feeding, take 1.1 lysate 50ul in the chain of streptomycin (800ug / ml) 5ml LB liquid medium, 150rpm 37 ℃ overnight culture.", "D22":"1.Cultivation Mhei-mcherry gene protein
•Measuring OD600 = 1.423, induced expression, take 1.2 full amount of liquid in the chain of streptomycin (800ug / ml) 30ml LB liquid medium, 150rpm 37 ℃ coated foil overnight culture.", "D23":"1.Extraction of cytoplasmic protein by ultrasonic crushing", "D24":"1.SDS page
•Check the results", "D27":"1.Extraction of cytoplasmic protein by ultrasonic crushing", "D28":"1.SDS page
•Check the results", "D29":"1.Enzyme activity detection
•UV spectrophotometer", "D30":"1.Enzyme activity detection
•Enzyme activity" }, "August":{ "D1":"1.Enzyme activity detection
•UV spectrophotometer", "D2":"1.Enzyme activity detection
•Enzyme activity", "D4":"1.Transformation opdA gene to DH5a competent Cells", "D5":"1.Cultivation opdA gene
•Sub-cultures were grown overnight to added suitable the concentration of antibiotics in 5ml LB and used to inoculate 30ml LB liquid to a starting OD600 of 0.5, cells were shaking culture at 37℃.", "D6":"1.Cultivation opdA gene
•When cultures reached a cell density of 1.0~2.0, used silver paper to make the culture be in dark ambient induce recombinant protein expression.", "D7":"1.Plasmid DNA extraction(8/3)
•Using Takara extraction kit", "D8":"1.Transformation opdA gene to DH5a competent Cells", "D9":"1.Cultivation opdA gene
•Sub-cultures were grown overnight to added suitable the concentration of antibiotics in 5ml LB and used to inoculate 30ml LB liquid to a starting OD600 of 0.5, cells were shaking culture at 37℃.", "D10":"1.Cultivation opdA gene
•When cultures reached a cell density of 1.0~2.0, used silver paper to make the culture be in dark ambient induce recombinant protein expression.", "D11":"1.Plasmid DNA extraction(8/3)
•Using Takara extraction kit", "D12":"1.Extraction of periplasmic and cytoplasmic protein
2.SDS page
•Check the results", "D15":"1.Extraction of periplasmic and cytoplasmic protein
2.SDS page
•Check the results", "D16":"1.Enzyme activity detection
•UV spectrophotometer", "D17":"1.Enzyme activity detection
•UV spectrophotometer", "D18":"1.Enzyme activity detection
•Enzyme activity", "D19":"1.Enzyme activity detection
•Enzyme activity", "D20":"1.HPLC
•The collection and preparation of samples", "D21":"1.HPLC
•The collection and preparation of samples", "D22":"1.Sample preparation
2.Standard working solution preparation", "D23":"1.HPLC", "D24":"1.HPLC", "D25":"1.Enzyme inhibition test", "D26":"1.Enzyme inhibition test", "D27":"1.Enzyme inhibition test", "D28":"1.Apoptosis test", "D29":"1.Apoptosis test", "D30":"1.Apoptosis test" }, "September":{ "D1":"1.OD 600 Reference point
•Add 100ul LUDOX 100% into wells A1，A2，A3，A4; add100ul of H2O into wells A2, B2, C2, D2. Measure absorbance at 600nm for all samples in standard measurement modes on plate reader.
2. Fluorescein fluorescence standard curve
•Spin down fluorescein stock tube to make sure pellet is at the bottom of tube. Prepare 2*fluorescein stock solution(100uM) by resuspending fluorescein in 1mL of 1xPBS.", "D2":"1.Interlab experiment
•Day 1:Transformed into Escherichia coil DH5a", "D3":"1.Interlab experiment
•Day 2: Picked 2 colonies from each plate and inoculated it into 5-10ml LB medium with Chloramphenicaol. Grew the cells overnight (16-18 hours) at 37℃and 220 rpm.", "D4":"1.Interlab experiment
•Day 3:Cell growth,sampling,and assay.", "D6":"1. OD 600 Reference point
•Add 100ul LUDOX 100% into wells A1，A2，A3，A4; add100ul of H2O into wells A2, B2, C2, D2. Measure absorbance at 600nm for all samples in standard measurement modes on plate reader.
2. Fluorescein fluorescence standard curve
•Spin down fluorescein stock tube to make sure pellet is at the bottom of tube. Prepare 2*fluorescein stock solution(100uM) by resuspending fluorescein in 1mL of 1xPBS.", "D7":"1.Interlab experiment
•Day 1:Transformed into Escherichia coil DH5a", "D8":"1.Interlab experiment
•Day 2: Picked 2 colonies from each plate and inoculated it into 5-10ml LB medium with Chloramphenicaol. Grew the cells overnight (16-18 hours) at 37℃and 220 rpm.", "D9":"1.Interlab experiment
•Day 3:Cell growth,sampling,and assay.", "D11":"1.PCR Bio-brick basic parts
Sample:
•pSB1C3 backbone
•Basic parts
2.Agarose gel
•Check the results", "D12":"1.Digestion
•Digestion the Bio-brick PCR product(9/11) with Dpn
•Digestion the backbone and plasmid with EcoRI and PstI overnight.
Sample:
•pSB1C3 backbone
•Basic parts", "D13":"1.Ligation
•Ligation DNA fragment(9/12) into the backbone plasmid pSB1C3.
2.Transformation
•Transformation of ligated product(9/12) to DH5a.", "D14":"1.Colony PCR
2.Agarose gel
•Check the insert gene fragment
3.Cultivation", "D15":"1.Plasmid DNA extraction(9/14)
•Using Takara extraction kit", "D16":"1.Digestion certification
•Digestion the plasmid (9/15) with EcoRI and PstI.
2.Agarose gel
•Check the insert gene fragment", "D20":"1.PCR Bio-brick basic parts
Sample:
•pSB1C3 backbone
•Composite parts
2.Agarose gel
•Check the results", "D21":"1.Digestion
•Digestion the Bio-brick PCR product(9/20) with DpnI
•Digestion the backbone and plasmid with EcoRI and PstI overnight.
Sample:
•pSB1C3 Terminator backbone
•Composite parts", "D22":"1.Ligation
•Ligation DNA fragment(9/22) into the plasmid backbone pSB1C3 Terminator.
2.Transformation
•Transformation of ligated product(9/22) to DH5a.", "D23":"1.Colony PCR
2.Agarose gel
•Check the insert gene fragment
3.Cultivation", "D24":"1.Plasmid DNA extraction(9/23)
•Using Takara extraction kit", "D25":"1.Digestion certification
•Digestion the plasmid (9/24) with EcoRI and PstI.
2.Agarose gel
•Check the insert gene fragment" }, "October":{ "D6":"1.PCR Bio-brick basic parts
Sample:
•pSB1C3 backbone
•Composite parts
2.Agarose gel
•Check the results", "D7":"1.Digestion
•Digestion the Bio-brick PCR product(10/6) with DpnI
•Digestion the backbone and plasmid with EcoRI and PstI overnight.
Sample:
•pSB1C3 Terminator backbone
•Composite parts", "D8":"1.Ligation
•Ligation DNA fragment(10/7) into the plasmid backbone pSB1C3 Terminator.
2.Transformation
•Transformation of ligated product(10/7) to DH5a.", "D9":"1.Colony PCR
2.Agarose gel
•Check the insert gene fragment
3.Cultivation", "D10":"1.Plasmid DNA extraction(10/9)
•Using Takara extraction kit", "D11":"1.Digestion certification
•Digestion the plasmid (10/10)with EcoRI and PstI.
2.Agarose gel
•Check the insert gene fragment" } };