Monday Feb. 1 Use sterile hoods Make sure autoclave is on and reserved for the class time Stock solutions: Prepare the following stock solutions, all dissolved in 50% ethanol. Make up the first solution in each table, then make up the following by serial dilution as shown in the table using 1.5 mL microcentrifuge tubes. Then aliquot 100 µL into 0.5 mL microcentrifuge tubes so that each group will have a set of the tubes of each auxin that they will need. (NAA and IAA made up fresh; protect IAA from light; concentrations 10,000X those given in the lab manual on pp. 32-33 and reproduced in tables below), and 1 mL of 50% ethanol as control (keep these by hoods) 1-NAA Required Add to Desired final stock concentrations solution 5 x 10-7 M 5 x 10-3 See #3 at 1.5 mL of M beginning 50% ethanol, of TA adjust to give prep correct concentration 3 x 10-7 M 3 x 10-3 0.84 mL 0.56 mL of M of 5 x 50% ethanol 3 M 1.5 x 10-7 M 1.5 x 0.7 mL of 0.7 mL of 1-NAA 3 M 3 x 10-3 50% ethanol M 0.5 x 10-7 M 0.5 x 0.47 mL 0.93 mL of 1-NAA 3 M of 1.5 x 50% ethanol 10-3 M 0 auxin 50% -- -- ethanol 2,4-D Required Add to Desired final concentrations solution 5 x 10-7 M 5 x 10-3 See #3 at 1.5 mL of 2,4-D M 2,4-D beginning 50% ethanol, of TA adjust to give prep correct concentration x 10-7 M x 10-3 0.84 mL 0.56 mL of 2,4-D M 2,4-D of 5 x 50% ethanol 3 M 1.5 x 10-7 M 1.5 x 0.7 mL of 0.7 mL of 2,4-D 3 M 3 x 10-3 50% ethanol M
Monday Feb. 1 Use sterile hoods Make sure autoclave is on and reserved for the class time Stock solutions: Prepare the following stock solutions, all dissolved in 50% ethanol. Make up the first solution in each table, then make up the following by serial dilution as shown in the table using 1.5 mL microcentrifuge tubes. Then aliquot 100 µL into 0.5 mL microcentrifuge tubes so that each group will have a set of the tubes of each auxin that they will need. (NAA and IAA made up fresh; protect IAA from light; concentrations 10,000X those given in the lab manual on pp. 32-33 and reproduced in tables below), and 1 mL of 50% ethanol as control (keep these by hoods) 1-NAA Desired final concentrations Required stock solution Add to 5 x 10-7 M 5 x 10-3 M See #3 at beginning of TA prep 1.5 mL of 50% ethanol, adjust to give correct concentration 3 x 10-7 M 3 x 10-3 M 0.84 mL of 5 x M 0.56 mL of 50% ethanol 1.5 x 10-7 M 1-NAA 1.5 x M 0.7 mL of 3 x 10-3 M 0.7 mL of 50% ethanol 0.5 x 10-7 M 1-NAA 0.5 x M 0.47 mL of 1.5 x 10-3 M 0.93 mL of 50% ethanol 0 auxin 50% ethanol -- -- 2,4-D Desired final concentrations Required solution Add to 5 x 10-7 M 2,4-D 5 x 10-3 M 2,4-D See #3 at beginning of TA prep 1.5 mL of 50% ethanol, adjust to give correct concentration 3 x 10-7 M 2,4-D 3 x 10-3 M 2,4-D 0.84 mL of 5 x M 0.56 mL of 50% ethanol 1.5 x 10-7 M 2,4-D 1.5 x M 0.7 mL of 3 x 10-3 M 0.7 mL of 50% ethanol 3 3 3
Villa-Cuesta%20and%20Hobbie%202016%20Genetic%20Lab_Instructor%20Guidelines_Arabidopsis.docx