Question

Directions: Please complete Prelecture Assignment and upload answers to Carmen as a PDF before coming to lecture. Before attempting this worksheet, please read Lab 8 as this worksheet accompanies the lab and helps you get your lab notebook ready by guiding you through the necessary calculations that will be needed to start the experiment. This lab takes more thought and effort to set-up, but faster to complete if you think about things now. Please bring a copy of the completed assignment to lecture and lab so that you will be ready to get started. Deadline: Posted on Carmen Understanding how to prepare stock solutions: 1. Given a 50 mM stock solution of sulfapyridine in DMSO, how will you prepare 100 HL of a 2 mM solution of this compound using modified Sauton's (SAU) medium to dilute the compound? Please show calculation below. (Hint: use MV-MV:) 50 MM sulfapyridine 1004 total volume atend Stock 2mM HL of 50 mM stock sulfapyridine solution + HL SAU-100 HL of a 2 mM sulfapyridine solution Understanding how to prepare the plate: 2. In this laboratory, you will be using a 96-well plate to conduct the experiment. It can be very confusing to know what goes in the wells of the plate if you haven't done this kind of experiment before. To help make this less confusing, as you read the "Prepare the plate" section in the lab manual, start at #1 and fill in the information to help you know what to add to each row/column of the 96-well plate. Please use the following abbreviations: commercial sulfanilamide (CSA), your synthesized sulfanilamide (mySA): sulfapyridine (SP), sulfadiazine (SD), sulfathiazole (ST), positive control (PC), and negative control (NC) to label the wells. Positive controls have maximal bacterial growth and negative controls have no bacteria. Controls do not have drugs present. 1 2 3 4 5 6 7 8 9 10 11 12 #1 L of distilled HO A X X X X X X X X _of 2% DMSOB UN Oooo L1 of_mmof_t0000000000003 Luik of_mmor_c0OO000000000 added to _mmof_t=0OOOOOOOO000 & towy 8:23 INN #4 Determine volume, []. & name of drug added to B7-B12 & rows C-G _mM of_TE ) 11. 01_mmof_ T Lid of distilled 140 a nonooo 700101 ᏙᏙᏙᏙᏙᏙᏙᏙᏙᏙᏙ Discard # Perform Serial Dilutions ul transferred #6 Add-HLE.coli to all wells EXCEPT #7 Which wells serve as your positive & negative controls ? __Controls __Controls

Answer 1

mi 8 final concentration M2 = N m, spyz0o 1 Stock solution M, = 50mm mv, - m₂ V2 V2 = 100w V = M₂ V2 M - 2 mM - Toox 2 - 4 Take 4ul and dilute it to 100g/ to get 2mM 3.A. V=500 (SAU) V2 2 500+50 CSA M = ? 200M M2: 26899) volume Initial volume - soul Final Concentration Initiat concentraction = 20 mm - -50+50-tooul - 10 mm at “Troy Final Concentration of esn 10 mm B. Initial Volume = soul, final volume = 100) Dilution = Find top 2 - Two fold ditution मन- c. V = 504 V = 1004 [50+50 M₂ = 1m 56X1 Son M = 10 mp4 M2 ?? V, - 50 pel V2 = 100k/ 150 +50] M, 2 10mm M₂=9 M₂ 2 59710 = 5 mM CSA in c] Final Volume = 100mling M=5 100 = Gout in 7,50 .6 25mite