Difference between revisions of "Team:CCU Taiwan/Notebook"
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    101 Competent Cells [DH5a]<br> |     101 Competent Cells [DH5a]<br> | ||
    Ligation pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18<br> |     Ligation pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18<br> | ||
| − | |||
| − | |||
    Digest pUCIDT_Lac1/ pUCIDT_Px16/ pUCIDT_Px18 with AgeI, EcoRI<br> |     Digest pUCIDT_Lac1/ pUCIDT_Px16/ pUCIDT_Px18 with AgeI, EcoRI<br> | ||
</p> | </p> | ||
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<p class="description">    Digestion – pGAPZ A-1+HIS4 cut with AgeI and EcoRI (x3)<br> | <p class="description">    Digestion – pGAPZ A-1+HIS4 cut with AgeI and EcoRI (x3)<br> | ||
    Ligation – pGAPZ A-1+HIS4 cut with AgeI and EcoRI (x3)<br><br> |     Ligation – pGAPZ A-1+HIS4 cut with AgeI and EcoRI (x3)<br><br> | ||
| − | + |     pGAPZ A_Px16, pGAPZ A_Px18 , pGAPZ A_Lac1 Traditional plasmid DNA extraction<br> | |
| + |     Yeast SMD1168 Transformation(pGAPZ A_Px16, pGAPZ A_Px18 , pGAPZ A_Lac1)<br> | ||
    Transform <br> |     Transform <br> | ||
      1. pGAPZ A-1 + HIS4 + Px16 <br> |       1. pGAPZ A-1 + HIS4 + Px16 <br> | ||
Revision as of 02:33, 18 October 2018
NOTEBOOK
Notebook
February
Molecular biology class
E. coli genomic DNA preparation
E. coli transformation
March
Instrument operation
April
Project brainstorming: Product Positioning, Monolignol proportion
May
Culture selection: compare Yeast, E. coli, Acetobacter aceti
Gene design
June
Interlab experiment: Calibration 1, 2, 3
July
7/3
Start Interlab experiment: cell measurement
7/5
Yeast Extract–Peptone–Dextrose (YPD) formulation
7/9
Yeast (X33) culture
7/13
Fundraising briefing session
7/16
Communicate with NCKU (Interlab & project)
7/18
Communicate with BIT (project)
August
8/5~8/12
Cell growth, sampling, and assay
Digest pGAPZ A(X3)/ pUCIDT_Lac1/ pUCIDT_Px16/ pUCIDT_Px18 with AgeI, EcoRI
Transformation pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18 (15µl) into 20µl ECOS™
101 Competent Cells [DH5a]
Ligation pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18
Digest pUCIDT_Lac1/ pUCIDT_Px16/ pUCIDT_Px18 with AgeI, EcoRI
8/13~8/19
Miniprep pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18
Digest pGAPZ A(X3)/ pUCIDT_Lac1/ pUCIDT_Px16/ pUCIDT_Px18 with AgeI, EcoRI
8/20~8/26
Gel extract
Digestion – pGAPZ A-1+HIS4 cut with AgeI and EcoRI (x3)
Ligation - pGAPZ A_Lac1/ pGAPZ A_Px16/ pGAPZ A_Px18
8/27~9/2
Digestion – pGAPZ A-1+HIS4 cut with AgeI and EcoRI (x3)
Ligation – pGAPZ A-1+HIS4 cut with AgeI and EcoRI (x3)
pGAPZ A_Px16, pGAPZ A_Px18 , pGAPZ A_Lac1 Traditional plasmid DNA extraction
Yeast SMD1168 Transformation(pGAPZ A_Px16, pGAPZ A_Px18 , pGAPZ A_Lac1)
Transform
1. pGAPZ A-1 + HIS4 + Px16
2. pGAPZ A-1 + HIS4 + Px18
3. pGAPZ A-1 + HIS4 + Lac1
Digest pGAPZ A HIS4(X2) with AgeI, EcoRI
September
9/3~9/9
PCR—pGAPZ A-1 + HIS4 + Px16, Px18, Lac1
miniprep – pGAPZ A-1 + HIS4 + Px16, Px18
Transform – pGAPZ A-1 + HIS4 + Lac1
Digestion – pGAPZ A-1 + HIS4+lac1
9/12~9/14
Colony PCR gel electrophoresis check - HIS4+Px16, HIS4+Lac1
Culture HIS4+Px16, HIS4+Lac1
Colony PCR gel electrophoresis check - HIS4+Px16, HIS4+Lac1
Yeast SMD1168 pGAPZ A_Px16, pGAPZ A_Px18, pGAPZ A_Lac1 colony PCR
Plasmid miniprep - HIS4+Lac1, HIS4+Px16
9/24~9/27
Culture HIS4+Lac1 x3
Prepare yeast competent cells (SMD1168)
Traditional plasmid DNA extraction Day1
Yeast SMD1168 pGAPZ A_Px16, pGAPZ A_Px18, pGAPZ A_Lac1 colony PCR
Yeast SMD1168 pGAPZ A_Px16, pGAPZ A_Px18, pGAPZ A_Lac1 Western blot
Prepare YPD plates
Prepare the amino acids of SD plates
Traditional plasmid DNA extraction Day2
Plasmids contain HIS4 cut with SalI
Yeast transformation - Px16, Px18, Lac1 (plasmids contain HIS4)
October
10/8~10/11
Prepare yeast competent cells (SMD1168)
Colony PCR (HIS4+Px16)
Safety (30℃, 50℃, 70℃)
Yeast transformation - Px16, Lac1 (plasmids contain HIS4)
Yeast SMD1168 pGAPZ A_Px16, pGAPZ A_Px18, pGAPZ A_Lac1 Western blot