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Unformatted text preview: 2 $HindlIland ^EcoRI Small DNA fragment (From freparation oflnsert) I 1tI ligase 2. 9 F!H2O 4 pl5 x ligationbuffer 4 1rl digested vector (200 ng) (From'?reparation of Vector') 2 pI HindIII and.EcoRI Large DNA fragment (From ?reparation oflnserf) I 1rI ligase 3. ll rrlHzO 4[d5xliguionbuffer 4 pl digested vestor (200 ng) (From *Preparation of Vector') I Fl ligase Tap to mir Place PCR tube into a 1.5 ml microcentrifuge tube and centrifuge for 15 seconds at 5,000 rpm. Incubarc l5-l6oc overnight (in the thermocycler, which in this case will serve as an expensive cooling bloct capable of mainAining a constant ' temperatre) . Store all the molecular weight marters, and your large and small DNA inserts inthe frws. . Empty'waste" container inthe trash caos. . Empty ice brrdcets ard return buckets to the shelf. . Wipe off1our bench with water and paper towels....
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This note was uploaded on 07/13/2008 for the course MCDB 104 taught by Professor Lee during the Summer '08 term at UCLA.
- Summer '08