JoVE
JoVE
教师资源中心
科研
行为学
生物化学
生物学
生物工程
癌症研究
化学
发育生物学
工程学
环境科学
遗传学
免疫与感染
医学
神经科学
JoVE 杂志
JoVE 实验百科全书
JoVE Chrome Extension
教育
生物学
化学
Clinical
工程学
环境科学
Pharmacology
物理学
心理学
统计学
JoVE 核
JoVE 科学教育
JoVE 实验室手册
JoVE Quiz
JoVE Business
Videos Mapped to your Course
发表
图书馆员
高中
关于
Sign-In
登录
联系我们
科研
JoVE 杂志
JoVE 实验百科全书
教育
JoVE 核
JoVE 科学教育
JoVE 实验室手册
高中
CN
EN - English
CN - 中文
DE - Deutsch
ES - Español
KR - 한국어
IT - Italiano
FR - Français
PT - Português
CN
EN - English
CN - 中文
DE - Deutsch
ES - Español
KR - 한국어
IT - Italiano
FR - Français
PT - Português
Close
科研
行为学
生物化学
生物工程
生物学
癌症研究
化学
发育生物学
工程学
环境科学
遗传学
免疫与感染
医学
神经科学
Products
JoVE 杂志
JoVE 实验百科全书
教育
生物学
化学
Clinical
工程学
环境科学
Pharmacology
物理学
心理学
统计学
Products
JoVE 核
JoVE 科学教育
JoVE 实验室手册
JoVE Quiz
JoVE Business
与您的课程匹配的视频资源
Teacher Resources
Get in Touch
Instant Trial
Log In
CN
EN - English
CN - 中文
DE - Deutsch
ES - Español
KR - 한국어
IT - Italiano
FR - Français
PT - Português
Journal
/
遗传学
/
A High-Throughput UAS-cDNA/ORF Plasmid Library Construction Using CRISPRmass
/
Large-Scale Transformation of UAS-cDNA/ORF Plasmids from CRISPR-Based Modular Assembly into
E. coli
JoVE 杂志
遗传学
需要订阅 J
o
VE 才能查看此.
登录或开始免费试用。
JoVE 杂志
遗传学
Large-Scale Transformation of UAS-cDNA/ORF Plasmids from CRISPR-Based Modular Assembly into
E. coli
Large-Scale Transformation of UAS-cDNA/ORF Plasmids from CRISPR-Based Modular Assembly into
E. coli
A High-Throughput UAS-cDNA/ORF Plasmid Library Construction Using CRISPRmass
DOI:
10.3791/201692-v
•
01:46 min
•
May 17, 2024
•
Si Xu*
1,2
,
Mengyu Chen*
1,2
,
Guang Chen
3
,
Si Luo
,
Wen Xue
2
,
Xuelian Liu
2
,
Jiwu Wang
2,3
,
Ping Wei
1
Clinical Research Institute, The Affiliated Nanhua Hospital, Hengyang Medical School
,
University of South China
,
2
Department of Neurology, The Affiliated Nanhua Hospital, Hengyang Medical School
,
University of South China
,
3
Department of Clinical Laboratory, The Affiliated Nanhua Hospital, Hengyang Medical School
,
University of South China
,
4
Shanghai Diabetes Institute, Shanghai Key Laboratory of Diabetes Mellitus, Shanghai Clinical Center for Diabetes
,
Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine
Tags
Transformation Of UAS-cDNA/ORF Plasmids From CRISPR-Based Modular Assembly Into E. Coli:1. Pre-chill 10 μL Tips And 1.5 ML Tubes In An Aluminum Cooling Block On Ice.2. Thaw Competent E. Coli Cells On Ice For 5 Minutes.3. Add 10 μL Of Competent Cells To Each Pre-chilled 1.5 ML Tube.4. Add 1 μL Of The UAS CDNA/ORF Plasmid Products To Each Tube.5. Gently Swirl The Tubes To Mix And Incubate On Ice For 30 Minutes.6. Heat Shock The Cells At 42°C For 1 Minute.7. Immediately Chill The Tubes On Ice For 2 Minutes.8. Add 100 μL Of Pre-warmed (37°C) SOC Medium To Each Tube.9. Incubate The Tubes In A Shaking Incubator At 250 RPM And 37°C For 1 Hour.10. Warm The LB Plates Containing The Appropriate Antibiotic To 37°C.11. Spread The Cells Onto The Warm Plates And Incubate At 37°C Overnight
Embed
添加到播放列表
Usage Statistics
Related Videos
Author Spotlight: Simplifying Genome-Wide Plasmid Library Construction Using CRISPRmass
Large-Scale Construction of UAS-cDNA/ORF Plasmids by CRISPR-Based Modular Assembly (CRISPRmass)
Large-Scale Transformation of UAS-cDNA/ORF Plasmids from CRISPR-Based Modular Assembly into
E. coli
Read Article