A lecture on classical Restriction Enzyme Cloning.
Time Stamps
0:26 — Primer Design
2:21 — How to download more base pairs from NCBI
4:05 — What should you order after designing?
5:32 — PCR needs two primers
7:22 — Sanger sequencing only needs one primer
8:25 — C/G Clamp
9:53 — Other considerations of primers
11:54 — It can flip directions
14:54 — Self ligation
16:45 — Calf intestinal phosphatase
19:08 — Why you might have to use one restriction enzyme
20:00 — Other types of cloning
21:06 — What goes in to the PCR reaction
22:15 — PCR amplicon
23:04 — Why cloning longer things are less efficient
25:31 — PCR Purification
28:15 — RE Digest
36:10 — Gel purification
38:51 — Kit gel purification
40:00 — Ligation
41:21 — Transformation
42:40 — Select bacteria
44:12 — Freezing
47:52 — Verification
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