Objectives: 09/04/2014 Lab Meeting Article Info Finish lab 3a
Discuss lab 3b Lab notebooks will be collected at the end of class
tomorrow
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Bellwork: 11/01/2013 1.330 mL = _________ L _______________ 2.
P-10/20/100/1000 measure what volumes?
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Today: 11/04/2013 1.How would you calculate the mass required
to make the following solutions: 5 mL of 2.5% glucose solution:
2000 mg = ___________g 5L = ___________mL 6.3cm = _________mm
Slide 4
Using the Spec: 1.Turn the instrument on and allow it to warm
up for 15 minutes. 2.Set your spec to the desired wavelength. 3.Use
a kim wipe to clean off any finger prints, oil, debris, etc., from
the outside of your test tube. 4.Insert the standard test tube
(deionized water) and blank the spec. 5.Insert your test sample and
read/record both %transmittance and absorbance. 6.Repeat steps 4
and 5 for each testable sample.
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A) Metric System IS: International system of measurement
PrefixSymbolFactor kilok1000 hectoh100 (base unit) no prefix----1
decid0.1 centic0.01 millim0.001 micro 0.000 001 (10 -6 ) nano n
0.000 000 001 (10 -9 )
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A) Metric System
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To easily convert between prefixes we can use the BS rule
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B) Measuring Volumes The main units we measure volumes in are
microliters, milliliters and liters 1000 L = 1 mL = 0.001 L *know
these conversions!!!!! Practice: 1) 125 mL = _________ L 2) 33 mL =
_________ L 3) 4 L = _________ mL 4) 200 L = _________ mL 125,000
0.033 4,000 0.200
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C) How do we measure volume in the lab? Graduated cylinder:
Measure more than 10 mL Pipet: measure between 0.5 mL &10 mL
Micropipet: less than 1 mL
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C) How do we measure volume in the lab? Given the following
volumes, what instrument would you use to measure that volume: 5)
100 L 6) 25 L 7) 0.005 mL 8) 56 mL 1) 100 mL 2) 25 mL 3) 5 mL 4) 10
mL Graduated Cylinder Graduated Cylinder Pipet Pipet or Graduated
Cylinder Micropipet
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D) How to use a pipet Draw this: In order to use a pipet you
much also have a pipet pump: Blue: 1- and 2-mL pipets Green: 5- and
10-mL pipets Red: 25- and 50-mL pipets
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Roll the gear so the top raises, this will draw up the liquid
into the pipet Roll the gear the opposite direction to expel the
liquid DO NOT push the top of the pipet pump this may break the
pump When selecting a pipet to use remember to use the smallest
pipet as possible to be as precise as possible
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E) How to use a micropipet Used to measure amounts less than 1
mL 3 sizes of pipets: P100.5 L to 10 L P100 10 L to 100 L P1000100
L to 1000 L Each micropipet has only a certain amount of precision
Make sure you pay attention to this!
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E) How to use a micropipet These are the readings for
micropipets Main difference between 20 and 10 is the top limit, it
still has the same unit places on the display
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E) How to use a micropipet P10/P20
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E) How to use a micropipet P100/P200
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E) How to use a micropipet P1000/P2000
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F) Making Solutions One of the most essential skills Labs often
buy the solid form of a compound because it is often cheaper and
many times multiple concentrations are used. Almost all reactions
involving proteins and nucleic acids occur in an aqueous solutions.
Solution contains 2 parts: Solute what is being dissolved Solvent
what us doing the dissolving
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F) Making Solutions To prepare a solution: 1. Measure out solid
mass of solute needed 2. Add solid to container with a little water
3. Dissolve solid 4. Add enough water to reach desired volume
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Bellwork: 09/23/2014 You need 50 L of D-Fructose for a
bioreactor. This D-Fructose must be at a concentration of 12 mg/mL.
How many kilograms of D-Fructose do you need?
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G) Concentrations A measure of how many particles are in a
certain volume Normality made be used for acids & bases only
Common Units of Concentration ConcentrationCommon Units of
Measurement mass/volumeg/L, mg/mL, g/mL, g/L % mass% MolarityM
(mol/L), mM (mmol/L), M (mol/L)
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H) Preparing Mass/Volume Concentration Mass/Volume
Concentration Equation Conc. desired x volume desired = mass of
solute (g/mL)(mL)(g) Example: How do you prepare 100 mL of 0.05
g/mL solution of hemoglobin? 1)Use the equation to determine mass
of solute. 0.05 g/mL x 100 mL = 5 g hemoglobin 2)Give directions
for making solution: Measure out 5 g of hemoglobin and pace into a
100 mL flask, add about 10-20 mL of dH 2 O. Dissolve all hemoglobin
then add enough water to make 100 mL of solution.
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H) Preparing Mass/Volume Concentration Mass/Volume
Concentration Equation Conc. desired x volume desired = mass of
solute (g/mL)(mL)(g) Practice: How do you prepare 150 mL of 2 mg/mL
gelatin? 1)Use the equation to determine mass of solute. 2 mg/mL x
150 mL = 300 mg gelatin = 0.3 g gelatin 2)Give directions for
making solution: Measure out 0.3 g of gelatin and dissolve in a
little bit of water. Add enough water to make 150 mL of
solution.
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% Mass/Volume Concentration Equation Convert % to decimal
Decimal % x volume desired = mass of solute (g/mL)(mL)(g) I)
Preparing % Mass/Volume Concentration Example: How do you prepare
50 mL of 10% NaCl solution? 1)Use the equation to determine mass of
solute. 0.10 g/mL x 50 mL = 5 g NaCl 2)Give directions for making
solution: Measure out 5 g of NaCl and dissolve in a small amount of
water. Add enough water to make 50 mL of solution. 10% 0.10
g/mL
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I) Preparing % Mass/Volume Concentration % Mass/Volume
Concentration Equation Convert % to decimal Decimal % x volume
desired = mass of solute (g/mL)(mL)(g) Example: How do you prepare
60 mL of 10% glucose? 1)Use the equation to determine mass of
solute. 0.10 g/mL x 60 mL = 6 g glucose 2)Give directions for
making solution: Measure out 6 g of glucose and dissolve in a small
amount of water. Add enough water to make 60 mL of solution. 10%
0.10 g/mL
Slide 26
09/17/2014 Your background must be complete prior to starting
the lab. Everyone must wear gloves, lab coat/apron, and eye
protection We are using Methylene Blue instead of Pyronin Y
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09/17/2014 Lab Changes We are using Methylene Blue instead of
Pyronin Y You will microwave your agarose to melt it. You will use
TAE buffer instead of TBE buffer Microfuge Tube Symbols:
Bromophenol Blue B Orange G G Methylene Blue M Safranin O O Xylene
Cyanol X Unknown ?
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General Critiques You need to know what you are doing before
you do it. Read your instructions and ask questions prior to
starting. Do your math first! Your peers are a very good resource,
dont follow me around asking for help when you have 20 peers that
can answer your question. Dont use the analytical balances unless
you need to! Remove scoops and close containers once your are done
using them
Slide 29
Power Supply Settings 100 V 50 minutes 35 amps Make sure the
wells are covered in TAE buffer Do not start the power supply until
four boxes are connected
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Bellwork: 09/19/2014 Get started! If you ran your gel
yesterday, work on entering your data into your lab notebook and
writing your conclusion. If you did not run your gel yesterday,
hustle to get the gel loaded and start the power supply. Once all
the gels are running we will start our mini presentations
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Bellwork: 09/22/2014 How would you prepare 10 mL of 50 mg/mL
CuSO 4 5H 2 O Formula: Total Volume x Concentration = mass of the
solute
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Lab 3e: Making Solutions of Differing Mass/Volume
Concentrations The regular lab report format is not needed for this
lab, but the following should be documented in your lab notebook:
Tables 3.8, 3.9, and 3.10 A print out of step h on page 47
Questions 6, 10, and 14 on page 49 Be sure to take two sets of spec
readings! PPE gloves, goggles, lab coat/apron
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Objectives: 09/26/2014 Complete through lab 3e and turn in your
lab notebook for grading Science article presentations Begin work
on Lab 3f
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Bellwork: 09/29/2014 1. If you were pouring a gel for DNA
agarose gel electrophoresis, where would you place the comb? Sketch
a picture and explain your reasoning. 2. Make sure the data you are
recording is actually your data, not an ideal from a recipe, and be
concise in your writing
Slide 35
Lab 3f Requirements The regular lab report format is not needed
for this lab, but the following should be documented in your lab
notebook: Tables 3.11, 3.12 Show your math for one tube in table
3.11 Run Part IV twice Once with the professionally made Biurets
and once with yours Do not do the spec/absorbance steps PPE gloves,
goggles, lab coat/apron We will not be doing labs 3g/h, but you are
responsible for knowing the math within those labs
Slide 36
Quick & Easy Bellwork: 1. How does gel electrophoresis
work? 2. Give an example of when a spectrophotometer would be used,
but the solution used to blank the device would not be water
Slide 37
J) Preparing solutions using Molarity Molarity Units: mol/L
mole = the number of particles in a substance that is equal to the
mass (in grams) on the periodic table Ex: 1 mole NaCl = _____ g 1
mol of any substance will not be the same as a different substance!
1 mole Na = 23.0 g1 mole Cl = 35.5 g 1 mole NaCl = 23.0 g + 35.5 g
= 58.5 g 58.5
Slide 38
J) Preparing solutions using Molarity Practice: Solve the
following. 1) 1 mol C 6 H 12 O 6 = ________ g 2) 1 mol CaCl 2 =
________ g 180.0g and 111.1g are called the molecular weight (MW)
or formula weight of its substance (12.0 x 6) + (1.0 x 12) + (16.0
x 6) (40.1 x 1) + (35.5 x 2) 180.0 111.1
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J) Preparing solutions using Molarity Molarity Concentration
Equation Volume wanted x molarity x MW = mass of solute
(L)(mol/L)(g/mol)(g) Example: How many grams are needed to make a
75 mL solution of 2M CaCl 2 ? 0.075 L x 2M x 111.1 g/mol = 16.67g
CaCl 2 Practice: How do you prepare 20 mL solution of 0.25 M NaCl?
0.020 L x 0.25M x 58.5 g/mol = 0.293 g NaCl Measure out 0.293g of
NaCl and dissolve in enough water to make 20 mL of solution.
Slide 40
What mass of calcium chloride (CaCl 2 ) is needed to make 125
mL of 0.55 M CaCl 2 solution?
Slide 41
1 st convert 125 mL into L (molarity = moles/liter) 125 mL =
0.125 L Then use the molarity concentration equation
Slide 42
Volume wanted x molarity x MW = mass of solute 0.125 L x 0.55 M
CaCl 2 x 111 grams/mole = 7.63 grams of CaCl 2 and add solvent up
to 125 mL
Slide 43
K) Dilutions Buying concentrated solutions can save money If
you need a very dilute solution starting from a more concentrated
solutions may be required
Slide 44
K) Dilutions Dilution Equation C 1 V 1 =C 2 V 2 C =
ConcentrationV = Volume 1 & 2 stand for different solutions
Concentrations can be in any unit as long as both are the same,
same with volume Example: How would you prepare 1 L of 1mg/mL
protein solution from 100mg/mL concentrated stock? C 1 = 1 mg/mLV 1
=1LC 2 =100 mg/mLV 2 =? (1 mg/mL)(1L) = (100 mg/mL)(V 2 )V 2 =10mL
Measure out 10 mL of protein stock solution and mix in enough water
to make 1 L of solution.
Slide 45
K) Dilutions Dilution Equation C 1 V 1 =C 2 V 2 C =
ConcentrationV = Volume 1 & 2 stand for different solutions
Concentrations can be in any unit as long as both are the same,
same with volume Practice: How do you make 200 mL of 10mM CaCl 2
from 75mM CaCl 2 solution? C 1 = 10 mMV 1 = 200 mLC 2 =75 mMV 2 =?
(10 mM)(200mL) = (75 mM)(V 2 )V 2 =26.6mL Measure out 26.6 mL of
CaCl 2 and mix in enough water to make 200 mL of solution.
Slide 46
Get to Work! Changes to the Lab: Step 3 (page 56) You do NOT
need to graph your data. The trend(s) should be obvious through a
data chart. For your conclusion: Possible errors Results with
evidence Practical use for this technique Thinking like a
Biotechnician, questions 1, 2, 3 A summary of page 58 why is pH
biologically important? Etc.
Slide 47
Get to Work! For your background section: Explain: mole,
molarity, Avogadro's number, and molecular weight Write out the
molarity concentration equation (include all units) Solve the
following, showing all of your work: What is the molarity of a
solution containing 9.478 grams of RuCl 3 in enough water to make
1.00 L of solution? How many grams of KMnO 4 are required to
prepare 1.0 L of a solution of 1.5 M KMnO 4 ?
Slide 48
Get to Work! For your background section: Explain: mole,
molarity, Avogadro's number, and molecular weight Write out the
molarity concentration equation (include all units) Solve the
following, showing all of your work: What is the molarity of a
solution containing 9.478 grams of RuCl 3 in enough water to make
1.00 L of solution? How many grams of KMnO 4 are required to
prepare 1.0 L of a solution of 1.5 M KMnO 4 ?
Slide 49
Unit Equations: Mass/Volume Concentration Equation Conc.
desired x volume desired = mass of solute (g/mL)(mL)(g) %
Mass/Volume Concentration Equation Convert % to decimal Decimal % x
volume desired = mass of solute (g/mL)(mL)(g)
Slide 50
Unit Equations: Molarity Concentration Equation Volume wanted x
molarity x MW = mass of solute (L)(mol/L)(g/mol)(g) Dilution
Equation C 1 V 1 =C 2 V 2 C = ConcentrationV = Volume 1 & 2
stand for different solutions Concentrations can be in any unit as
long as both are the same, same with volume