Cloning with Restriction Enzymes (5 Levels) – Traditional Molecular Cloning

References/Resources: https://www.patreon.com/the_Crux

Playlist – Genetic Engineering: https://www.youtube.com/playlist?list=PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf

Molecular Cloning with restriction enzymes can be very easy or very challenging. This video discusses the fundamental considerations of traditional restriction enzyme based molecular cloning. We discuss sticky and blunt ends, the process of blunting, and how all of this (among other things) can be used to understand the different levels of molecular cloning. Doesn’t matter if you are doing subcloning or a PCR based cloning, you will eventually find yourself in one of the levels.

Jump to your favorite section:
00:00 Cloning and Tetris
00:56 Sticky ends
01:18 Where do you cut?
02:29 Blunt ends
02:49 Cheating with Klenow
04:42 Level 1
07:04 Level 2
08:21 Level 3
11:57 Level 4
14:25 Level 5

Patreon Page: https://www.patreon.com/the_Crux

Suggested videos:
Plasmids (pUC19, pBR322, etc.): https://youtu.be/2pAM-FTFI2c
Yeast Cloning Vectors: https://youtu.be/SHE0lpMiitA
BACs/Fosmids: https://youtu.be/Yktq7d3WDVs
Lambda/Cosmids: https://youtu.be/SHE0lpMiitA

*Must watch playlists*
All videos on Genetic Engineering: https://www.youtube.com/playlist?list=PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf
All videos on DNA Replication: https://youtube.com/playlist?list=PL0Ymnd-zt4IgLefDaeqicJbCBnbLqnDJ7
All videos on Transcription: https://www.youtube.com/playlist?list=PL0Ymnd-zt4Ij2VcAOHNUweftSElVsc-MX
All videos on Translation: https://www.youtube.com/playlist?list=PL0Ymnd-zt4IjAAAMlPLDIXbs692Yp14xa

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