Difference between revisions of "Team:CCU Taiwan/Notebook"
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Revision as of 09:42, 16 October 2018
EXPERIMENTS
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)
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)
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
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
Plasmid miniprep - HIS4+Lac1, HIS4+Px16
9/24~9/27
Culture HIS4+Lac1 x3
Prepare yeast competent cells (SMD1168)
Traditional plasmid DNA extraction Day1
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)
10/8~10/11
Prepare yeast competent cells (SMD1168)
Colony PCR (HIS4+Px16)
Safety (30℃, 50℃, 70℃)
Yeast transformation - Px16, Lac1 (plasmids contain HIS4)
Safety end