Team:USTC/Notebook/8 28

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Date: 8.28

Recorder: Yu Xie Plasmid Extraction for pSB1C3 containing sfGFP1-10

Recorder: Yu Xie PCR of bacteria which have recombinant plasmid pSB1C3 containing sfGFP1-10

Procedure:

1.Prepare 4 PCR tubes and sequentially add：

sample 1,2,3,4
Sterilized ddH2O 9 μL
2×HiFi-PCR Master 12.5 μL
Template(bacteria) 1.5 μL
Primer 1 1 μL
Primer 2 1 μL
total 25 μL

2.PCR reaction Parameters setting：

stage temperature time
Pre-Duration 95 10 min
Duration 95 1 min
Anneal 58 1 min
Extend 72 1 min 50s
Post-Extend 72 10 min
Final 4 --

30 cycles(Duration ~ Extend)

Agarose gel electrophoresis Results: 图片名称 succeed!

Recorder: Yu Xie PCR of recombinant plasmid pSB1C3 containing sfGFP1-10

Procedure:

  1. Prepare 4 PCR tubes and sequentially add：
sample 1,2,3,4
Sterilized ddH2O 9.5 μL
2×HiFi-PCR Master 12.5 μL
Template(plasmid) 1 μL
Primer 1 1 μL
Primer 2 1 μL
total 25 μL
  1. PCR reaction Parameters setting：
stage temperature time
Pre-Duration 95 10 min
Duration 95 1 min
Anneal 58 1 min
Extend 72 1 min 15s
Post-Extend 72 10 min
Final 4 --

30 cycles(Duration ~ Extend)

Agarose gel electrophoresis Results: 图片名称 succeed!

Date: 8.29

Colony PCR to examine ADW Recorder: Yinchenguang Lyu

6.25 μL 2×Taq PCR Master Mix 2 μL culture containing Ecoli 0.4 μL bio-p 0.4 μL M13-f 3.45 μL ddH2O

Double Digest to examine ADW and C3AD35 Recorder: Yinchenguang Lyu

result(along with that of the former part)

a (from left to right: Trans 2K plus 2(contain Gelred), digested-ADW1, digested-ADW2, C3AD35(1), C3AD35(2), ADW1, ADW2, ADW3, ADW4, ADW5, ADW6)

Date：8.30

Plasmid extraction of pGAL1(site mutation) Recorder: Xuefeng Meng

sample 1 2 3 4
concentratin(ng/μL) 379.6 232.9 277.2 375.4
A260/A280 1.81 1.85 1.90 1.90
A260/A230 1.42 1.72 2.33 2.31