General Course Information

Biology 275LCell and Molecular Biology

TA information

• Name: Jesse Owens• Email: jbowens@hawaii.edu• For Office Hours PLEASE Email Me and I will

set up a time to meet with you

• Secondary TA: Steffanie Snyder

Laboratory Notebook

• You are required to keep a binder containing:

– The lab manual– notes, handouts and corrected lab summaries

returned to you

Grading

10 Lab Summaries (20 Points)10 Lab Quizzes (5 Points)2 Practical Exams (75 Points)2 Written Exams (75 Points)

Total 550 Points

Laboratory Summaries • Must include the following sections:– Purpose 25%– Materials and Methods 15%– Data 25%– Conclusion 35%

• More will be discussed at the end of class

LATE ASSIGNMENTS

• Summaries are due at the start of the lab period the week after the lab experiments

• Absolutely NO LATE WORK WILL BE ACCEPTED• (policy for all labs – not my idea)

Lab Quizzes• At the beginning of each lab section you will

be taking a quiz (should last 10 minutes)• The Quizzes will test you on:– Introductions to the Current Lab– Methods and Materials– CalculationsMAKE SURE YOU READ AND UNDERSTAND THE NEW

LAB BEFORE YOU COME IN

• You must come on time to take the quiz. NO LATE QUIZZES WILL BE GIVEN

Lab Examinations• Two 1-hour lab examinations• Two Practical Examination (more will be

discussed at a later time) – Exam #1– Exam #2CONCEPTS WILL BE CUMMULATIVE

• Refer to your lab manual for the exact dates

Return of Assignments• Assignments will be returned to you during the

scheduled lab periods

Biology WebpageBiology 275L web page

www.biology.hawaii.edu/275l

Username: UH Login Password: UH Password

You can find the: • Lab Manual• Individual Lab section webpages

Laboratory Attendance• Attendance is mandatory• Except under extenuating circumstances, there

will be NO OPPORTUNITY for you to attend another section

• If you cannot attend your lab section, you must notify your TA (me) in advance

• You will not be allowed into another section without proper permission

• The other TA wont let you in unless I give them the head’s up

Laboratory Safety• You will be working with many toxic and

carcinogenic substances in lab• Commit to memory the following– Location of the MSDS folder– Fire extinguisher– Eye Wash– Important Numbers (emergency: 66911)– Chemical Shower (outside)– First Aid kit

THINGS TO BRING EVERY CLASS

– CLOSED TOED SHOES– Safety Goggles/Glasses– Lab Coat– Gloves (FORGOT ASK LAB PARTNER)– Blue/Black Pen– Text and Notebook– Lab Summary– Calculator

Laboratory Safety

• Abide by the following rules at all times:– No smoking, drinking, or eating– Wash your hands before and after each lab– Always wear covered shoes, gloves, safety goggles and a

lab coat while performing experiments– Dispose of chemicals/microbes in the proper containers– Wipe your bench with disinfectant before and after each

lab period

Laboratory Safety Cont.– Broken glassware, slides and coverslips must be

disposed of into the “GLASS ONLY” boxes– Know how to use the Material Safety Data Sheets

(MSDS folder) found in the back of the room– Report any injuries to your TA immediately– Remove all gloves and lab coats before you leave the

room– Don’t throw food or drinks in the trash, throw them

away outside• These guidelines will help to protect you from

the harmful materials used in lab

Fire Alarm Rules (VIII)• When the fire alarm sounds, prepare to leave

the room immediately• Your assistant TA will help anyone with

disabilities to leave the room first• Your TA will make sure everyone has left the

room• Meet at Varney’s Circle so your TA can make

sure everyone is okay

Laboratory Equipment• You will be using expensive precision equipment during

the semester• These instruments are delicate and you will be instructed

on their use and proper care• If you have questions regarding the use and care of the

equipment don’t hesitate to ask questions• Notify your TA immediately if a piece of equipment is

not working properly so that it can be repaired• If an accident happens tell me right away, you wont get

in trouble

Cleaning Up• If you do not clean up after yourself, one point

will be deducted from your lab summary• At the end of the lab period be sure to:– Check your lab manual for special “cleaning up”

instructions– Put back items you used from your drawer– Clean and return all supplies to their proper places– Place used glassware and plastics in the appropriate

containers– Wipe your bench with disinfectant

Labs on Holidays (IX)• Re-scheduling lab sections that fall on

holidays is extremely difficult to do• The Biology Program asks for your

cooperation in attending labs that fall on holidays

• If for some reason you are unable to attend lab, notify your TA as soon as possible

TA Evaluations• To enable the Biology Program to

continually improve lab courses, we need your input

• You will have two opportunities to evaluate your TA– 1 x during the mid semester– End of the semester

Lab 0: Introductory Lab

Biology 275LCell and Molecular Biology

Micropipettes• Most molecular biological experiments require very

small volumes (microliters) of reagents

– 1L = 1000 ml = 1,000,000 l = 1x106 l– 1 ml = 0.001 L = 1000 l = 1x103 l– 1 l = 0.001 ml = 0.000001 L = 1x10-6 L

• Micropipettes are special precision instruments designed to deliver volumes ranging from 0.1 l to 1000 l

Pipetman MicropipettePlunger buttonPlunger button

Volume adjusterVolume adjusterTip ejector buttonTip ejector button

Pipette shaft/barrelPipette shaft/barrel

Volume indicatorVolume indicator

Ejector armEjector arm

Plastic tipPlastic tip

Pipette bodyPipette body

Sizes of Micropipettes

MicropipetMicropipet Volume RangeVolume Range

P-1000P-1000

P-100P-100

P-10P-10

100-1000 100-1000 ll10-10010-100ll1-101-10ll

Reading Volumes

1 1 1

0 = 1000 l 0 = 100 l 0 = 10 l

0 0 0

P-1000 P-100 P-10Never go above or below the designated volumes

Use of Micropipettes• To draw up a solution– Rotate the volume adjuster to the

desired volume– Place the appropriate tip on the barrel– Press the plunger to the first stop– Insert the tip into the solution– Draw up the solution by gradually

releasing the plunger– Check to see that there are no air

bubbles present in the tip

Use of Micropipettes Cont.• To expel the solution

– Touch the tip against the side of the tube or container into which the solution will be expelled

– Press the plunger to the second stop to make sure everything is expelled

– Remove the pipette tip from the container with the plunger still pressed

– Release the plunger slowly– Eject tip into the appropriate waste

container by pressing the tip-ejection button

Care of Micropipettes• The following rules should be observed at all times:

– Never place a pipette near the edge of the bench where it can get knocked over

– Never rotate the adjuster below or beyond the designated ranges

– Never immerse the pipette barrel in liquid– Never lay the pipette down or hold it horizontally when

there is liquid in the tip– Always wipe the pipette barrel when you are done– Always use the appropriate tip for each pipette

Sterile Tips• Do not touch the tips with your bare hands• Keep tip boxes closed as much as possible• Always change tips when changing solutions• Dispose of tips into the “Pipette Tips Discard”

container unless otherwise specified

Centrifuges• Always balance the tubes to prevent damage to

the rotor• Make sure top is screwed on before spinning• Lengths and speeds of centrifuges– Flash spins– Timed spins– 1000-14,000 rpm• What is Supernatant and Pellet?

Aseptic Technique• Microbes are ubiquitous (they go everywhere)• Reduces the chances of contaminating yourself

or the experiment• Involves the following:– Sterilizing all media and equipment– Keeping containers closed as much as possible– Disinfecting your hands and bench frequently

Laboratory Exercises• Practice using the micropipettes– Try pipetting 5l– Try .9 ml

• Review C1V1 = C2V2

– For some it may be easier to remember the formula as

CInitialVInitial = CFinalVFinal

• Sample Question

Lab Summaries

• Turned into your TA at the beginning of class (none for this week, only concentration questions)

• NO Late Assignments are accepted• Must be typed, however it is recommended that

graphs be done by hand • Must Include Purpose, Method, Results/Data, and

Conclusion. • An example Lab Summary will be given to you next

week

GradesIf you want to guarantee do well:

• don't skip anything

• write clearly, logically, and concise

• read the labmanual before coming to class and understand why we are doing the experiments that were doing

• (do these I'll guarantee you an A, this isn't hard but you can't be lazy)

Laboratory Summaries

PurposeShort paragraph stating the objective of the experiment.

It is worth 25%

Laboratory SummariesMaterials and Methods

Provides all pertinent information needed to repeat experiment. Not the details of the protocol.

• Organism• Sample preparation techniques• Reagents• Quantities• Temperatures• Times• Instrumentation• Statistical Method

It is worth 15%

Laboratory SummariesResults (DATA)

Explain what you saw but don’t draw any conclusions yet. You’ll want to include everything that you record and also diagrams, graphs and tables in this section.

Data: please refer to Appendix C for formatting.It is Worth 25%

Laboratory Summaries

Discussion The discussion section should explain all data, tables, & graphs

presented in the results section and your interpretations of those results in your own words.

Write short, simple, and pertinent sentences. No fluff, this is science and I’ll take off points if you write too many useless words

Summary inclusion questions should be answered separately and turned in on a separate sheet.

It is worth 35%

Laboratory Summaries

The weekly lab summaries should be no more than 2 spaced typed pages

(excluding the results section)I STOP READING AFTER 2

PAGES!

Anthrax!

• How many students are infected?• What % of students is that? Total/#infected• Who is the Dirty Host?

Mutants

.00000000000000000000001% Good

Real Mutant

• 99.999999999999999999999% Not Good

Before you leave today…

• Sign waiver• Think about who you want to sit with• Remember to do lab questions for next week• Remember to read over manual so that you

will do great on next week’s quiz