Dilution Calculation and Preparation
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Questions and Answers

What is the formula for calculating dilution factor?

IC / FC = Dilution factor

What is the purpose of performing serial dilutions in microbiology?

To reduce the concentration of the sample for accurate colony counting.

If you wish to dilute a 15% stock solution to a final concentration of 5%, what is the dilution factor?

15/5 = 3 (dilution factor)

Explain the significance of the dilution factor in microbiological serial dilutions.

<p>The dilution factor is the reciprocal of the dilution and is used to calculate the concentration of Colony Forming Units (CFU) per milliliter.</p> Signup and view all the answers

What is the purpose of using aseptic techniques in microbiology?

<p>To prevent contamination from pathogens and minimize the risk of infection.</p> Signup and view all the answers

How can the dilution factor be calculated from the dilution label (e.g., 10-1, 10-2, etc.)?

<p>The dilution factor is 10 raised to the power of the exponent in the dilution label.</p> Signup and view all the answers

List three procedures in microbiology where aseptic technique is commonly used.

<p>Transferring cultures, Inoculating media, Isolation of pure culture</p> Signup and view all the answers

What formula is used to calculate the CFU/ml from the number of colonies on a plate and the dilution factor?

<p>CFU/ml = Number of colonies on plate x Reciprocal of dilution factor</p> Signup and view all the answers

What are some sources of contamination that proper aseptic techniques aim to prevent?

<p>Airborne microorganisms, Microbes from researcher’s body/lab coat, Microbes in dust</p> Signup and view all the answers

Explain the importance of maintaining a 'Pure stock culture' in microbiology.

<p>To ensure the accuracy and reliability of experimental results.</p> Signup and view all the answers

How is the unit of CFU/ml interpreted in microbiology?

<p>CFU/ml represents Colony Forming Units per milliliter, indicating the concentration of viable cells in the sample.</p> Signup and view all the answers

Why is it essential to count plates with colony numbers between 30 to 300 in microbiological analysis?

<p>To ensure the accuracy of the colony count and statistical reliability of the results.</p> Signup and view all the answers

What is the formula for calculating the final volume of a solution after dilution?

<p>C1V1=C2V2</p> Signup and view all the answers

How does the total amount of a compound (moles or mass) change when diluting a solution?

<p>It does not change.</p> Signup and view all the answers

In a solution, how is the total mass calculated?

<p>Concentration x volume = total mass</p> Signup and view all the answers

What is the equivalent weight of NaCl if its molecular weight is 58 g/mol?

<p>NaCl = 2.9 g</p> Signup and view all the answers

How can a 1X saline sodium citrate (SSC) solution be prepared from a 20X stock SSC solution?

<p>By diluting one unit (volume) of 20X solution with 19 volumes of water.</p> Signup and view all the answers

What is the formula for dilution factor when diluting a solution?

<p>CiVi=CfVf</p> Signup and view all the answers

Study Notes

Solution Preparation

  • v/v = volume of reagent per final solution volume
  • Simple dilution: combining a unit volume of a liquid material with an appropriate volume of a solvent liquid to achieve the desired concentration
  • Example: preparing 1X SSC solution from 20X stock SSC solution by diluting one unit of 20X solution with 19 volumes of water

Complex Solution

  • To make a complex solution with two or more solid solutes, treat each solute individually
  • Calculate the mass of each solute needed based on its molecular weight (MW)
  • Example: preparing 5 liters of 50mM NaCl, 10mM Tris-Cl solution

Dilution Formula

  • C1V1 = C2V2, where C1 and C2 are initial and final concentrations, and V1 and V2 are initial and final volumes
  • Concentration x volume = total mass
  • Example: 50mg of X in 100ml diluted to 1L still has 50mg of X

Serial Dilution

  • Methods for diluting a solution, depending on experimental requirements
  • Determine the dilution factor before diluting a solution
  • Steps for serial dilution: take several sterile tubes, label with dilution factors, add diluent and stock sample, mix well, and repeat

Plate Count

  • Procedure for viable counting using serial dilutions of the sample and the pour-plate method
  • Calculation: No. of colonies on plate x reciprocal of dilution of sample = CFU / ml
  • Example: 32 colonies on plate of 1:10,000 dilution = 320,000 CFU/mL

Colony Counting System

  • Count plates with colony numbers between 30 and 300
  • Use formula to calculate: Cells (CFU) per milliliter of original sample = Number of colonies in plate count x Dilution Factor / Volume of sample taken

Aseptic Techniques

  • Definition: practices and procedures to prevent contamination from pathogens
  • Importance: to maintain pure stock culture while transferring to a new media
  • Types of contamination: airborne microorganisms, microbes from researcher's body/lab coat, lab surfaces, dust, and unsterilized glassware & equipment
  • Proper aseptic techniques can minimize and eliminate the risk of contamination

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Description

Learn about diluting solutions with examples like preparing a 1X saline sodium citrate solution from a 20X stock solution. Understand the process for making complex solutions with multiple solid solutes.

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