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Plasmid Isolation (Alkaline Lysis) - G-Biosciences

G-Biosciences 1-800-628-7730 1-314-991-6034 A Geno Technology, Inc. (USA) brand name think proteins! think G-Biosciences PR047 Plasmid Isolation ( alkaline lysis ) Teacher s Guidebook (Cat. # BE-310) Page 2 of 12 MATERIALS INCLUDED .. 3 SPECIAL HANDLING INSTRUCTIONS .. 3 ADDITIONAL EQUIPMENT REQUIRED .. 3 TIME REQUIRED .. 3 OBJECTIVES .. 4 BACKGROUND .. 4 TEACHER S PRE EXPERIMENT SET UP .. 5 PREPARATION OF LB BROTH .. 5 PREPARE BACTERIAL STOCKS .. 5 5 VISUALIZATION OF PURIFIED PLASMIDS .. 6 MATERIALS FOR EACH GROUP .. 7 PROCEDURE .. 7 RESULTS, ANALYSIS & ASSESSMENT .. 9 MATERIALS INCLUDED This kit has enough materials and reagents for 24 students (six groups of four students). 1 vial Bac: Plasmid 1 Agar stab 1 vial LB Broth 2 bottles LB Broth 1 vial AMP (Ampicillin) 1 vial Sterile Water 1 bottle Cell Suspension Solution 1 bottle lysis buffer 1 bottle Neutralization buffer 1 vial TE buffer 1 bottle Precipitation Solution 50 Centrifuge Tubes ( ) SPECIAL HANDLING INSTRUCTIONS Store Plasmid 1 Agar Stab at 4 C Store AMP (Ampicillin) frozen until ready to us

Add 200 µ l Lysis Buffer and mix the contents by gently inverting the tube 4-5 times. This solution contains sodium hydroxide and SDS (sodium dodecyl sulfate). The sodium hydroxide denatures the plasmid and chromosomal DNA into single strands. SDS, an ionic (charged) detergent dissolv es the phospholipids in the membrane

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