BIOL309 Study Guide - Final Guide: Protein Tag, Microtome, Microrna
27 Jan 2017
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Document Summary
Used to determine the direction of insert of a cloned fragment for which a restriction map is known. Can perform digestions and determine the orientation of the insert based on the expected sizes. Must also consider whether you are preparing a linear or circular map. Double digests: cut with two enzymes at the same time to determine orientation. Probes can be designed to bind specific sequences or group of sequences with a region in common (gene family members) There are numerous methods for labeling probes and method chosen depends on: type of template available, whether a strand-specific probe is needed, whether a very sensitive (highly-labeled) probe is needed. Incubating membrane with denature probe for sequence of interest. Before you start screening: probe design depends on what you have available: other sequences, templates for probes, antibodies, you must anticipate how many sequences might bind your probe so you can design the hybridization conditions.
