recursal/Fanatic-Fandom huggingface dataset:
Gram Stain Grading

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## Steven's Class

### Materials

\*\*\*\*\*\*\*Re-plate *Bacillus* stock a few days or a week before to check stock quality (have had repeated performance problems)

* 6 NA plates: *Bacillus subtilis*
* 6 **Nutrient broths**: *Serratia marcescens*
* Gram stain materials
* Sterile H2O tubes

## Original

### Materials

* 7 **broths** of *Proteus vulgaris* @ 30 Degrees
* 7 **PLATES** of *Bacillus subtilis* @ 30 Degrees
* Gram Stain Supplies
Radio Daze

"**Radio Daze**" is the tenth episode in the fourth season of *Punky Brewster*.

## Summary

Punky's favorite rock radio station went bankrupt and is now acquired by a firm that performs drama readings. In order to encourage kids' interest in older radio, the new owners sponsor a contest where whichever group gives the best performance will get a trip to Disneyland. Punky, Cherie and Marguax learn about old-time radio and how the DJs made their own sound effects right in the studio, which looks like fun, but Punky's zeal is tempered when she must take on Garth Goobler (Jason Hervey) and Grant Goobler (Ryan Rushton), the sons of a famous radio star. Henry encourages Punky to get to work on a script, telling her she can never become an astronaut or like profession if she only worries about competitors. The Gooblers have a strong script, and they steal a box of sound effect equipment from Punky and Cherie, but the girls quickly gather some more sound effect equipment and are able to pull off the win.

## Cast

* Soleil Moon Frye as Punky Brewster
* Cherie Johnson as Cherie Johnson
* George Gaynes as Henry Warnimont
* Susie Garrett as Betty Johnson
* Ami Foster as Margaux Kramer
* Sandy as Brandon "the Wonder Dog"
* Jason Hervey as Garth Goobler
* Ryan Rushton as Grant Goobler
Antibiotic Sensitivity

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### **Period 1**

***Materials***

* 1 broth culture of *Staphylococcus aureus /* table x 2 classes = 12-14

* 1 NB culture of *Escheria coli /* table x 2 classes = 12-14

* Sterile dry swabs, 4 swabs / per pair x 2 classes = ~52

* Antibiotic Sensitivity Discs and Dispenser

* Nutrient agar plates (2 per pair)

### **Period 2**

* Plates from period 1 (24/48 hrs in incubator?)
General Semester Preparations - Prep Lab and More

## General things that should get taken care of before each semester

* Deep cleaning of all prep lab cupboards
* Throw away all unused items from the previous semester
	+ All unused sterile broth media can be discarded.
	+ Unused sterile plates can be bagged & saved for Micro 60 Aseptic technique lab. Unused sterile nutrient agar plates can be used in Bio 10.
* Lab schedule on prep lab doors and exterior windows 313 & 317
* Clean and organize all benches
* Deep clean glassware
* Deep clean and recheck all slides
* Vacuum bottom drawers of student lab benches
* Vacuum microscope cupboards

ADD MORE!!!
Punky P.I.

**"Punky P.I."** was a segment on *It's Punky Brewster* that aired on the sixth episode during Season 2.

## Synopsis

Punky launches a private investigation firm at their school.

## Trivia

* This segment's title is a play on the TV police drama series title *Magnum P.I.*
* Herbert's likeness bears close resemblance to Avery Bigelow from the "Pretty Ugly" episode in Season 1. Avery also attends the same school Punky and his friends go to.
* This segment was paired with a repeat of "Fish Story" from Season 1.
Bio 16 Photosynthesis Lab

### To prepare all phenol red:

* .025g/1L DI H2O
* Adjust pH to JUST turn red with 1M NaOH - Add one drop at a time!

## Photosynthesis

### Each Bench

* 125mL Erlenmyer flask
* 75 mL of Phenol Red
	+ =75mL x # benches x # classes = 75 mL x 6 benches x 2 classes = 900mL
* Straw
* 3 screw-cap tubes
* tube rack
* 2 x 3" pieces of Elodea
* Funnel for tubesa
* Setup grow rack in 313 to have an area for the tube racks
* piece of aluminum foil (to cover tube)
* "Used Elodea" dish
* tweezers to remove elodea from tube

## Respiration

### Per bench

* 3 Beakers with:
	+ 1 x germinating seeds (~20 seeds)
	+ 1 empty
	+ 1x same amount of non germinating seeds (~20)

* flask of phenol red =~200mL
* 200mL phenol red x 6 benches x 2 classes = 2400mL
* "Used Seeds" dish

### Per bench

* 3 pieces of chalk
* 1 sandwhich bag
* 1 empty beaker
* 1 beaker with 50 mL of vinegar
* Balances on back bench
* scoops
Bio 100 Properties of Water

**Exercise 1 – Polarity of Water**

**Exercise 2 – Adhesion and Cohesion**

**Exercise 3 – Capillary Action**

**Exercise 4 – Solubility**

**Exercise 5 – Surface Tension**

**Exercise 6 – Density of Water (Instructor demonstration)**

 1000mL beaker of room temp water with white paper back

5mL manual pipette with tip

flask of cold red water in ice chest

blue ice cube (in Freezer)

oil cube in freezer

small beaker of oil
Nutrient Broth Tubes

## **Making Nutrient Broth Tubes**

Each 16 x 125 tube = 6 mL of Nutrient Broth (NB)

### **Ingredients**:

* Nutrient Broth Powder (found in media cupboard)
* D.I. Water

### **Supplies**

* 1000mL beaker
* 500mL graduated cylinder
* large stir bar
* Hot plate / stir plate
* Balance
* Weigh boat
* Repeat pipettor with tip

### **Directions**

1. Measure the appropriate amount of NB powder in a weigh boat

on the balance

1. Add NB powder to flask as per directions on the container
	1. # of test tubes x 6mL each tube = Total mL NB needed (usually

round up to the next 100 mL, it is ok to have a little extra)

1. 1. Total mL NB / 1000 mL NB (in the directions) = fraction of

powder  from directions you will need

1. 1. 1. For example: If you are making 600mL of NB, you will

600mL/1000mL= 0.6

1. 1. 1. Then: The directions calls for 8g of powder for 1000mL of

Broth so…. 8g powder x 0.6 of the recipe = 4.8 g of powder to make 600mL of N

1. 1. 1. * Or ... (600mL NB /1000mL NB for recipe) = (x g powder needed / 8 g

powder for recipe)

1. 1. 1. * + 8 g powder for recipe (600mL NB /1000mL NB for recipe) = x g

powder needed

1. 1. 1. * + 8 (600/1000) = x
				+ 8 x 0.6 = x
				+ 4.8 = x (powder needed)
				+ Add stir bar

1. Turn heat to ~100 degrees and stir until broth is clear and

well mixed

1. Use Repeat pipettor to pipette 6mL into each tube
2. Cap each tube with plastic caps
3. Add a small piece of autoclave tape to top of tube rack
4. Autoclave for 15-17 minutes
Chalkley's Solution

> BACK TO CULTURING PROTISTS
>
>

MAKES 10 L of a 1x Solution

(The ingredients describe a 10x solution, must be diluted to
1x for use with protists)

3. Store the 1x solution in a media bottle until ready to culture or subculture your protists

### **Variations**:

* **For paramecium:** To prepare the media:Dilute Chalkley's to 1x solution with spring water ( I actually use just tap water). Add one alfalfa (I found that pressed alfalfa pellets work nicely) to 1L 1x boiling Chalkley’s Solution. To Culture: Add 15 boiled wheat seeds to 1 L 1x cooled alfalfa Chalkley's solution. Inoculate with 150mL culture. Culture will peak in 3 weeks. To Subculture by adding 200 mL fresh medium and inoculate with 5-6 mL established culture. Maintenance: Subculture every 3 to 4 weeks?

* **For Amoeba:** To Prepare the Media: Dilute Chalkley's Solution to 1 x with boiled spring water. Let cool. To Culture: Add 2 rice or wheat grains for every 50mL of 1x solution. To subculture: If more than 50 Amoeba are visible at 1 field of view in 4x, then it is time to subculture. Split in half. Maintenance: Media should be changed every month.

* **For Euglena:** To prepare the media: Dilute Chalkley's to 1x. To culture: For ever 1 L 1x Chalkley's, dilute 3x with spring water, add 20 wheat or rice grains and 5mL or 1 tsp of powdered milk. Then boil for 5-10 minutes. Let stand uncovered for 24 hours. Subculture: Shake and add to smaller container. Inoculate with 50-100mL culture. This is smelly. The population can crash within weeks, if there is a decrease, inoculate to new media.

* Paramecia (Alfalfa Chalkley’s Solution)
* Euglena
* Amoeba
Bio 10 Genetics Lab Set-Up

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## Notes

There is very little to this lab, as of yet. The students can work together to finish the lab manual genetics

problems.

Some instructors may opt to use the computers for Genscope. If they do, check with them to see if they need copies or if they need the computers put out.
MacConkey Lab

## Pre-Lab

Materials: Give fecal test tubes to each student

## Lab 1

1 MacConkey Agar Plate / Student

	+ Autoclave 1 empty tube per student (for students to aliquot for Methyl Red Test)
* SIM Agar Deep:
	+ SIM Medium, 16 x 100 mm Tube, 8 ml Deep
	+ Read here
* Urea broth
	+ Pre-sterilize water and tubes but this broth cannot be heated
	+ make with sterile technique
	+ Prep sterile objects the day before and prep broth just before class
	+ 2mL broth tubes - more broth will make a slower reaction time
* TSI Agar Slants
	+ Pull top 150 tubes with new rack @ 12mL
	+ Inoculating Needles
	+ Note: Bigger Butts Please!

## Lab 3

The incubated tests from Lab 2 = Mr-VP / Simmons / SIM / Urea / TSI Tubes = 1 each / student

For MR-VP:

* Sterile Transfer Pipettes = 1/ student
* Sterile Culture tube (methyl red test) = 1/student
* 3 Methyl Red Dropper Bottles
* 3 x Barritt's Reagent A
* 3 x Barritt's Reagent B

For SIM:

* 3 x Kovac's Reagent

#### Reagent amounts

* MR-VP:
	+ Methyl Red: 1mL broth : 3 drops Methyl Red
	+ Barritt's Reagent A: 4 drops
	+ Barritt's Reagent B: 2 drops
* SIMS:
	+ Kovac's: 5 drops
Allen Who?

**"Allen Who?"** was the first segment on the twelfth episode in Season 2 of the cartoon spinoff series *It's Punky Brewster*.

## Trivia

* This is the only episode in which Henry mentioned Glomer's name, as he was interested to know who he is. Punky covered up the situation by mentioning that Glomer is Allen's surname.
* This segment was paired with a repeat of "Spell-bound".
MICRO 5 Aseptic Technique

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#### **Materials Su2022 (closer to lab manual):**

* NA slants or plates of *Serratia marcescens* = 1/bench (Making tubes for Tuesday's Streak Plate Lab)
* NA slants or plates of *Bacillus subtilis* = 1/bench (Making tubes for Tuesday's Streak Plate Lab)
* Nutrient broths = 1 / stu +2 sterile nutrient broths for instructor demo
* NA plates (leftovers?) = 1/ student

#### **Materials Su2021:**

* NA slants of *Serratia marcescens* = 1/bench
* NA slants of *Bacillus subtilis* = 1/bench
* Nutrient broths = 1 / stu +2 sterile nutrient broths for instructor demo
Bio 16 3-Second Rule

#### As Lab Manual States Pg 8 - S22

* 36 NA plates = 6 per group
* 6 sterile petri dishes = 1/ group
* forceps = 1/ group
* 1 box of alcohol prep pads (to disinfect forceps)
* sharpies
* Canned fruit (enough for 6 pieces per group)

#### Steven's Class

* 40 NA plates
* 10 empty sterile petri dishes
* 10 forceps
* 1 box of alcohol prep pads (to disinfect forceps)
* sharpies
* Steven will supply the canned fruit

#### **Steven's Class**

* Incubated Plates
* 40 NA plates
* 10 empty sterile petri dishes
* 10 forceps
* 1 box of alcohol prep pads (to disinfect forceps)
* sharpies
* Steven will supply the canned fruit

## Lab 3

### Steven's Class Only

* Incubated Plates
* Sharpies
Henry Falls in Love, part 1

"**Henry Falls in Love, part 1**" is the fifteenth episode in the first season of *Punky Brewster*.

## Synopsis

Henry is reunited with a woman he planned to marry in 1941; when Pearl Harbor was attacked, he enlisted in the Merchant Marines and did not see her again until now.

## Cast

* Soleil Moon Frye as Punky Brewster
* George Gaynes as Henry Warnimont
* Susie Garrett as Betty Johnson
* Cherie Johnson as Cherie Johnson
* Ami Foster as Margaux Kramer
* Casey Ellison as Allen Anderson
* Sandy as Brandon "the Wonder Dog"
* Allyn Ann McLerie as Maggie McLerie
* Robin Bach as Jeffries
Planaria

## About

   Planarians are cute little worm like critters that have tiny eyespots on them when viewed under a microscope. They are very simple to care for.

 Quick Facts

 **Class** | Rhabditophora

* **Diet:** Carnivore (and scavenger)
* **Lifespan:** Potentially 3 years
* **Temp:** 72-75 Fahrenheit
* **Habitat:** Freshwater

## Feeding

1. Pull out the liver from the freezer. When doing this for the first time, please ask someone to show it to you. It is typically in a container with a white lid.
2. Grab a scapel and open the liver container.
3. Cut out a very small piece of the liver with the scapel. You can cut a few tiny pieces for them.
4. Take out the pieces with some forceps if you need to, or just use the scapel.
5. Put the pieces in the bottom of the planaria tank.
6. You are done!
7. Be sure to clean the scalpel after use.

*\*\*Planaria should be fed roughly once a week!\*\**

## Cleaning

Planaria need fairly clean water to survive. Here is how to maintain their tank.

* Clean out about 25% to 50% of the water one day after feeding the critters.
* Pull out any remaining white liver chunks during this cleaning.
* Use the turkey baster to pick up the liver pieces.
* Be careful to not harm the planaria!
* Check your cleaning apparatus to make sure no planaria got stuck in it.

## Links & Resources

Here are some interesting and useful links about planarians.

* Basic Care
* Simple care guide with pictures
* Cool planaria facts!
* An entire website dedicated to planarians
GMO Instructor Notes

## LAB 1

**Locations of things!**

* Ice is in the chest in the refrigerator
* The InstaGene **TEST** and  **NON-GMO** tubes are in the ice chest
* The **+ CONTROL tubes** are in the freezer

* The **Master Mix** and **Primer** tubes are in the freezer

* The **PCR** tubes are on the benches

**What else to do**:

1. Turn on Water bath to 95 Degrees
2. Master Mix and Primers
3. Get the master mix and primers from freezer and thaw

1. Add 7ul **G** PRIMER **to** GMM-MAS **(master mix)tube**

and mix

1. Aliquot 65ul of **GMM-MAS** to each  of the 5 red **GMM** tube and put on ice
2. Add 7ul PPRIMER **to** PMM-MAS **(master mix) tube and mix**
3. Aliquot 65ul of **PMM-MAS** to each of the 5 green **PMM** tube and put on ice

Please return the primers to the freezer, thank you!!!

## LAB 2

**Locations of things**

* The Student Samples and MWR’s (ladders) are in the freezer
* The ice is in the fridge
* The gels are on the back bench

**PCR Student Samples:**

* The Loading Dye has already been added to the PCR student samples and MWR tubes

* Each bench gets 1 tube of Molecular Weight Ruler “MWR” and their student sample

* Students will load 20ul of each sample and MWR into the gels

**Agarose Gels:**

* I made 1 extra gel in case of mistake
* I have molded the gels but the students can remove the tape and the combs, then load them into the boxes with the TAE buffer by your instruction.

**Run Time**

* The gels can be run at 120V for 30 to 40 minutes, or when you see the orange loading gel ¾ to the end

**Staining and** Unstaining

* I have provided gloves for the person transferring the dyed gel to the rinsing bucket

* Please remind the students to not dump the FastBlast down the sink, they can return it to the bottle when finished with staining

* The 40-55 degree instruction is just about as hot as the water gets – be careful, a little hot

* Rinse by the instructions, but have them keep rinsing until they can see the bands
Micro 5 Labs

***WORK IN PROGRESS: just putting everything here to make it easier!***

For all labs, make sure there is a
copy of the atlas in lab that is referred to repeatedly in the manual.

* None

* Loops

* Tubes

* Petri dishes with agar

* Metal petri plate sleeves

* Incubator

* petri plate sleeve

* sterile swabs

* sharpies in each desk

* incubator

* Prepared slides of paramecium (30)

* Kimwipes

* Lens paper

* *rhizopus*
* prepared slides of:

*(30 each)*

* Plate Cultures and broth cultures of *Bacillus subtilis* and *Staph aureus*

* loops

* slides

* staining trays with slide racks bibulous paper

* lens paper

* crystal

violet stain

* lens cleaner

* immersion oil

* squeezy water bottles

* Vaseline

* Loops

* slides

* staining trays

* bibulous paper

* immersion oil

* crystal violet

* Gram’s Iodine

* 95% ethanol and safranin

* squeezy water bottles

* loops

* pH strips

* slides

* milk

* yogurt

* incubator

* loops

* yogurt

* incubator

* Weighing paper

* balances

* 14 media bottles and caps

* nutrient agar powder

* 7 graduated cylinders (100ml)

* hot water bath?  They use stove

* empty petri plates (70)

* 7 metal sleeves

* balances

* 1 ml sterile pipettes (14)

* manual pipettors for 1 ml

pipettes

* 56 4.5 ml tubes of 0.85% NaCl

* 28 AIA agar plates

* alcohol beakers

* glass spreaders (7)

* vortexer

* forceps (14)

* loops

* dissecting scopes

* 65 Penassay Agar plates

* loops

* nutrient broth cultures of bacteria (names

not specified?)

**Effectiveness of Different**
Disinfectants in controlling microbial growth

nutrient agar plates (2 per student)

sterile swabs (2 per student)

microban

10% bleach

Lysol wipes

70% ethanol

**Intro to clinical micro**

None

**Indigenous Microflora**

Per student:

1 BAP

2 sterile swabs

gram stain supplies

microscope supplies

**Evaluating Antibiotics**

broth culture of *Staph aureus and E. coli,* one per student: nutrient agar plate and sterile  swabs, antibiotic impregnated disks

**Use of the micropipettor**

P1000, p200, p20 micropipettors at least 12 of each, tips for all, micro tubes

**Into to PCR**

PCR maching, p20 micropipettorswith tips (1 per pair), ice bucket (one per pair), one per student: paper cup, transfer pipette, colored microtube, insta gene mix.

Microcentrifuge (1) 2 water baths with different temps, 0.9% saline solution

Electrophoreses boxes (12) and power supply (3), TAE running buffer, loading dye, staining
tray, 1% agarose gels (enough to make one gel per pair of students)

**Transformation of Bacterial Cells**
with Plasmid DNA

competent E coli cells, pUC 18, LB broth, 12 LB plates p100 p200 micropipettors and tips,
alcohol, spreaders
Millipede Care

## Introduction

On July 21, 2016 we adopted three millipedes. They are being kept and cared for in the Room 313 terrarium.

A large, docile terrestrial animal, also known as the thousand-legger. This millipede lives a long time; is easy to care for; and has many segments, each bearing 2 pairs of appendages. Each millipede is approximately 3 to 4” long.

Read more about millipedes by clicking on the links at the bottom of the page.

**Currently, we do not have any millipedes.**

## Quick Facts

 **Class** | Diplopoda

* **Diet:** Detritivore
* **Lifespan:** 2-5 years
* **Temp:** ~25 Degrees Celsius
* **Habitat:** Terrestrial, typically forest floor dwellers
* **Humidity:** High
* **Activity Level:** Nocturnal

## Care

These little guys are some of the easiest insects to care for. They require little attention, however, here are some things that will keep them satisfied.

1. They require a moist environment, so mist the terrarium one to four times per week, check it daily to make sure that it is moist but avoiding molds. This also helps the plants in the terrarium.
2. Diet: always have rotting leaves and forest bits in the terrarium for them to eat, but we can also provide them with fish flakes, raw potato, apple, lettuce, mushrooms, carrots. If they are rotting, even better!
	* Chitin is required for their exoskeleton - oak leaves are great for this.
3. Remove any food that develops mold. Increase ventilation if mold becomes a problem.

## Links:

* Wikipedia information on millipedes

* Millipede Care Information

* Millipede Pet Care Information

* Another pet info page

* Carolina Arthropod Care Instructions

* Carolina Millipede Care Sheet
Food Contamination

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## Materials

### Day 1

**Hamburger / ground beef / fresh spinach Portion :: *He will do only the solid food for this lab***

***Materials: (per pair of students or 2 per bench)***

* 1 x sterile forceps or 1 sterile spatula
* manual pipettor
* 2 x sterile 0.1 - 1.0 pipette tips
* 1 x 9mL dilution blank
* 2 x 99 mL dilution blank
* 6 x sterile petri dishes
* 1 x 250 mL bottle of autoclaved and melted NA in a hot water bath @ 45 degrees Celcius (not too hot!)
	+ Actually, 1 per bench (6-8 bottles)
* lab tape and sharpies

Other materials

* Balances
* Extra disposal tub for pipette tips
* new weighing dishes (clean and still in sleeve)
* vortexers ( have 2 for the class)
* Water bath(s) @ 45 degrees
* Some extra sterile spatulas
Aquatic Animal Care Schedule

This is an outline of how to care for the aquatic animals in the lab. Specific details on how to do each task can be found at each tanks' page.

## **MONDAY**

*CHECK WATER LEVELS FOR ALL*

* ***Coral Tank***

1. Check filter sponges, protein skimmer, and back of tank, clean if needed
2. Check salt levels if time permits
3. Spot clean tanks
4. Add algae remover

* ***Touch Tank***

1. Check filter sponge, clean if needed
2. Check salt levels if time permits
3. Spot clean tanks

* ***Saltwater Tank***

1. Open filter, make sure it looks fine, rinse off white bag if needed
2. Check salt levels if time permits
3. Spot clean if needed

* ***Goldfish***

1. Clean tank with magnet sponge if needed
2. Feed

* ***Guppies***

1. Look under lid holding pots to make sure no guppies are trapped
2. Feed

## TUESDAY

*CHECK WATER LEVELS FOR ALL*

*Look over tanks, make sure everything looks good and functional*

* ***Coral Tank***

1. Check on animals

* ***Touch Tank***

1. Check on animals

* ***Saltwater Tank***

1. Check on animals

* ***Goldfish***

1. Feed

* ***Guppies***

1. Feed

## WEDNESDAY

*CHECK WATER LEVELS FOR ALL*

* ***Coral Tank***

1. Check filter sponges and protein skimmer, clean if needed
2. Check salt levels if time permits
3. Spot clean tanks

* ***Touch Tank***

1. Check filter sponge, clean if needed
2. Check salt levels if time permits
3. Spot clean tanks

* ***Saltwater Tank***

1. Check salt levels if time permits
2. Spot clean if needed

* ***Goldfish***

1. Feed

* ***Guppies***

1. Feed
2. Clean or do water change if needed

## THURSDAY

*CHECK WATER LEVELS FOR ALL*

* ***Coral Tank***

1. Check bioballs in back of tank, clean if needed
2. Check salt levels, if needed

* ***Touch Tank***

1. Do a water change, if needed
2. Check salt levels, if needed (or if water change is done)

* ***Saltwater Tank***

1. Check on animals
2. Check salt levels if time permits

* ***Goldfish***

1. Feed

* ***Guppies***

1. Feed

## FRIDAY

*CHECK WATER LEVELS FOR ALL*

* ***Coral Tank***

1. Feed all animals
2. Clean filters for weekend
3. Add algae remover

* ***Touch Tank***

1. Feed all animals
2. Clean filters for weekend

* ***Saltwater Tank***

1. Feed all animals
2. Check on animals

* ***Goldfish***

1. Feed all animals
2. Clean tank with magnet sponge

* ***Guppies***

1. Feed all animals
2. Make sure no guppies are trapped!
Coliform Analysis

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1. 2/student x Regular strength (1x) lactose broth + upside down Durham tube. (12mL) in a 16x150mm tt with plastic cap.
2. 1/student x Double strength (2x) lactose broth + upside down Durham tube. (10mL) in a 20x150mm tt with metal cap.
3. Sterile 10 mL pipettes + pipetters
4. Student water samples

### Methods

**Regular Strength Lactose Broth (# of students x 2 x 12mL = amount to make)**

* 3.0g Beef Extract

* 5.0g Peptone
* 5.0g Lactose
* 1000mL media DI Water
* Adjust to pH 6.8-7.0

*Can use combination of 0.2M KH 2 PO 4 & 0.2M Na 2 HPO 4 . Final concentration of buffer should fall into 0.02M to 0.1M. Put 1/10 th into DI H2O for total we need.*
**Double Strength Lactose Broth (# of students x 10mL = amount to make)**

* 6.0g Beef Extract
* 10.0g Peptone
* 10.0g Lactose
* 1000mL media DI Water.

## Lab 2

### Materials

* Brilliant green bile broth. 15 mL/ tt and 1 tt per student. 16x150mm tt w/ duram tube and a metal cap.
* Inoculating loop

*# of students x 15mL = amount to make. Premixed powder in media cabinet.*

## LAB 3

### Materials

1. EMB plates 1 per student plus 2 plates for Lab coordinator to streak controls.
2. NB cultures of *Escherichia coli* & *Eneterobacter aerogenes* for lab cooridinator to streak. FOR CONTROLS
Concepts Prep Instructions

#### Instructions

\*Prepare 6 large (~2cm) cubes and 6 small (~1cm) cubes for each class section + some extras for good measure. I have been making about 800-1000mL for 9 classes. MAkes 4-5 flats to cut up. This will leave you with a little extra just in case.

#### Preparation Materials (@1L)

* 10 mL Bromeresol Purple .4% (0.4g powder + 100mL H2O)
* 20g agar powder
* 890 mL DI H2O
* 100mL .01M NaOH

#### Methods

1. In 500mL media bottle, add 20g of agar powder
2. Add 890mL H2O
3. To make cubes, melt the agar,
4. Add 100mL .01M NaOH, and 10 mL Bromocresol Purple
5. Stir until dissolved
6. Autoclave and store
7. To make cubes, put the bottle in a warm bath to melt the agar
8. Once melted, mix gently and pour into square container
9. Let sit or refrigerate until solid
10. Remove from container
11. Cut 2 cm and 1 cm cubes (do the best you can).

#### **Setup Materials Per station (Have 2 stations)**

* 1 x 25% vinegar
* 1 x spoon
* 1-2 x calculators
* 2 x rulers
* 1 dish with 1 x 1cm cube and 1 x 2cm cube (have 1 dish of cubes per each group = 6 / class)
* 1 x small soaking dish
* 1- 2 x discard beakers or flask and beaker for used vinegar and used cubes

### **Exercise 2:**

#### **About**

   This is an amazing experiment that covers the scientific method. This is for Exercise 2.

#### Instructions

Prepare  ***3 SETS*** of the experiment. There will be 3 stations in each lab for this one single exercise.

#### Materials and Methods

Prepare 18 x 500mL flasks with 200mL nutrient broth in each. You only need 15 but I always make a couple extra just in case of contamination. Follow the instructions on the bottle but...

#### **Nutrient Broth**

(These are instructions for 1 L of broth. I usually fix up 3600mL for about 18 flasks)

1. Suspend 8 g of nutrient broth powder in 1L of DI water.
2. Mix thoroughly on a hot plate
3. Boil for 1 minute to dissolve the powder
4. Aloquot 200mL into individual 500 mL flasks
5. Autoclave all flasks with stoppers on liquid for 15 minutes (121 degrees)
6. All stoppers and tubes that will be used must be autoclaved too
7. Immediately put appropriate stoppers and tubes on the flasks after autoclaving

#### **Flasks**

* A: Inoculate 3 open flasks with yucky swabs from yucky places and incubate
* B: Pour 3 into new, un‐autoclaved flasks and close
* C: Keep 3 Sealed completely with #7 stoppers, do not open after leaving the autoclave
* D: Affix 3 with straight glass tubes before autoclaving
* E: Affix 3 with hook tubes before autoclaving
Pringsheim's Soil Water

> BACK TO CULTURING PROTISTS
>
>

This is a media for culturing protists: Volvox and Spirogyra (Green Algae).

Makes about ~1.5L of Solution

3. Fill three-fourths full with distilled or deionized water and then add one gram of calcium carbonate (CaCO3).
4. Loosely plug or cap the jar and heat the mixture on a hot plate until it steams. You can use a pressure cooker but not an autoclave.... hmmm...
5. Allow the mixture to steam for about one hour and then cool to room temperature.
6. Repeat this procedure for two to three consecutive days.
7. The soil-water should be as close to sterile as it can get.

### Variations:

**Euglena** Preparations: Before steaming, add one-fourth of a softened pea seed, without the seed coat, to enhance growth. Culturing: 1L Solution, add20 wheat grains and t tsp of powdered milk and boil for 5-10 minutes. Dilute x3 and allow to cool for 24 hours.

**Spirogyra**: Preparations: Do not add CaCO3.

**Volvox**: Preparations: Make sure it is at pH7. Subculture: Volvox needs to bee subcultured often.
Heat Sensitivity Lab

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## Summer 2021 (Steven's Class)

* 30 NA plates
* 6 broths each: *Staphylococcus aureus, Escerichia coli, Bacillus cereus,* and *Bacillus subtilis*.
	+ 72 hours old
* 4 x Water baths and / or hot plates (40, 60, 80 & 100oC)
* 8 thermometers
* test tube tongs.

##### PM class (Spring 2024)

* 6 each broth cultures: Sa, Ec, Bc, Bs (~72hrs)
* Each table does one temp – **25**, 40, 60, 80, 100
* 2 plates per table – each in half (4 microbes) = 2x6 = 12 plates

**Disinfectant lab**

* 24 sterile swabs
* Each table does 2 of the 4 chemicals
* 4 Na/ table - 4x6 (Clean plate and dirty plate for each chem) = 24 NA

## It is a test of bacterial resistance to heat.

We will turn on the hot plates as needed so that we can control the rise in temperature of the water in the beakers.

We will sample the cultures at 40, 60, 80, and 100 oC.
All in Henry's Family

**"All in Henry's Family"** was the only original segment on the fourth episode of the second season of *It's Punky Brewster*.

## Synopsis

Henry pays a visit to his unusual sisters in the country and takes Punky, Cherie and Brandon with him.

## Trivia

* This is the only segment in the series in which Allen Anderson and Margaux Kramer do not appear.
* This segment's title is a play on the sitcom title *All in the Family*.
* Unlike the other original segments in Season 2, this segment appears to be the only one animated using traditional cel animation - a practice which was being phased out around the time this season aired.
* This segment was paired with a rerun of the Season 1 segment "Double Your Punky".
Bio 10 Enzyme Lab Note on Improvements

## Anything you would like to add.... Ideas? Things that do not work well? Let me know!!

I would like to test this lab with turnips, however with the tests I have done so far, it is far less consistent. I am presently doing some research to find a certain buffer that helps the process. Turnips are easy to prepare and they are not smelly! Much more pleasant. We will see in the coming (S2016) semester, what I find.

Danielle
Streak Plate

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* Broth Tubes: *Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Klebsiella pneumoniae* = 6 each
* ~~Empty Sterile broth tubes = one per student - for instructor to mix broths~~
* Sterile transfer pipettes for instructor

### Lab 2 - RESULTS

* Incubated Plates
* Gram stain materials

## Streak Plate Practice

### Lab 1

* 2 NA plates / student
* BROTH: 3 x *E. coli* , 3 x *K. pneumoniae* / per class and 6 *x S. aureus* and 6 x *B. subtilis* / class.

### Lab2

* Incubated Plates
* 2 NA plates / student
* BROTH: 3 x *E. coli* , 3 x *K. pneumoniae* / per class and 6 *x S. aureus* and 6 x *B. subtilis* / class.

## Streak Plate Grading - Updated F22

### Lab 1

* 6 broths each: *Staphylococcus aureus* and *Escherichia coli*
* 1 NA plate per student

## Original (Lab Manual) p. 67

Using Koch's Postulates TSA+l and yogurt for streak plate practice

## Original Instructions

### Lab 1 - DO 2 TIMES - Prep for Streak Plate 1 and Streak Plate 2

Per pair

* 1 x broths *Serratia marcescens* (Made in yesterday's Aseptic Lab)
* 1 x broths *Bacillus subtilis* (Made in yesterday's Aseptic Lab)
* 2 x NA plate

### Lab 2

Per pair

* Incubated Plates (See results and use for new cultures)
* 2 x NA plates to re-streak
* Sterile H2O
Caught in the Act

**"Caught in the Act"** was the first segment of the thirteenth episode and series finale of the cartoon spinoff series *It's Punky Brewster*.

## Summary

Just before an important photographic assignment, Glomer gives Henry amnesia after he discovers him in the kitchen and confronts him about it.

## Trivia

* It is unknown what repeat segment this episode was paired with.
Bio 16 5 Understanding Science - Second Rule

(per section, **double**):

	+ Same materials as usual, it would be helpful if we could have 3 cans of peaches (1 for each pair of tables to help avoid cross-contamination and bottle necks with setup) If we can’t have 3 cans, we’ll make it work!

#### Steven's Class

* Pasteur's flasks without labels
* 40 NA plates
* 10 empty sterile petri dishes
* 10 forceps
* 1 box of alcohol prep pads (to disinfect forceps)
* sharpies
* Steven will supply the canned fruit

#### **Steven's Class**

* Incubated Plates
* 40 NA plates
* 10 empty sterile petri dishes
* 10 forceps
* 1 box of alcohol prep pads (to disinfect forceps)
* sharpies
* Steven will supply the canned fruit

## Lab 3

### Steven's Class Only

* Incubated Plates
* Sharpies
Micro 5 CA-MRSA Lab

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## **(Steven's Class)\*\*\*\*\*\***

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**\*\*\*\*Make sure you are following the correct version\*\*\*\***

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### Lab 1

(Per Student)

* 1 sterile dry swab
* 1 MSA plate
* 1 NA plate

### Lab 2

(Per Student)

* 1 MSA plate

### Lab 3

Per student

* 1 LB plate
* 1 LB-amp plate
* gram stain
* H2O2 bottles

### Lab 4

Per student

* observe plates (no materials)

## Original (Lab Manual Version)

### Lab 1

* 2 x NA plates
* 2 x MSA plates

### Lab 2

* Incubate Plates
* 1 x MSA plate
* *S. aureus* control = 1/ bench

### Lab 3

* Incubated Plates
* Catalase
* LB
* LB + amp
* Blood Agar
* Gram Stain
* Sterile H2O
Gram Stain with Unknown

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* *Staphylococcus aureus* (*24 hours)*
* *Bacillus subtilis* *(24 hours)*
* *Escherichia coli (24 hours)*

(48-72hrs)

* *Staphylococcus epidermidis* *(48-72hrs)*
* *Pseudomonas aeruginosa* *(48-72 hrs)*
* *Corynebacterium xerosis**(48-72 hrs)*

### **Additional Information**

I do this as an unknown for the students to test their own technique.  I would like *,* and perhaps one more (I will go through the plates again to see what is available).  I do not need one set of these per student. Two sets for the entire class will be enough. - Steven
Nitrate Reduction Test

Directions for Hardy Diagnostics Reagents A, B, and C

Method of Use: Heavily
inoculate Nitrate Broth (Cat. no. K42) or Nitrate Agar with the test organism.
When using Nitrate Agar, inoculate both the slant and butt of medium. Incubate
24-48 hours at 35ºC. To determine nitrite production, **add 5 drops of Nitrate Reagent A and 5 drops Nitrate Reagent B to the**
medium**. Shake gently to mix the reagents. Examine for the appearance of a**
deep red color within 1-2 minutes. If the result is negative, i.e. no color
development, **confirm the negative**
finding by adding Nitrate Reagent C (approximately 6.0mg or 0.006g) **to the**
tube containing the previously added Nitrate Reagents A and B. Examine for the
appearance of a deep red color within 5-10 minutes.

If using nitrate
reagents for commercial identification strips, such as API ®, consult the manufacturer's literature.

=== **INTERPRETATION**
OF RESULTS **===**
A positive nitrite
reduction is denoted by the appearance of a deep red color change after the
addition of Nitrate Reagents A and B. Lack of color development denotes a **presumptive**negative
nitrite reduction test. Development of a red color following the addition of
Nitrate Reagent C (zinc dust), confirms the negative nitrite reduction test
obtained in the first phase of the test. Lack of color development after the
addition of zinc dust indicates that the nitrate was reduced beyond the nitrite
reaction to nitrogen gas and constitutes a positive nitrate reduction reaction.
Remember When

"**Remember When**" is the 21st episode in the third season of *Punky Brewster*.

## Summary

After a long day of collecting snowballs for storage and storing them in the refrigerator's freezer, Punky, Cherie, Margaux, and Brandon are ready to wind down. The worst blizzard in years strikes Chicago, however, and a series of mishaps leaves Henry, Betty, Punky, Cherie, Margaux, and Brandon trapped in the apartment without power or heat, with conditions too severe to travel in. With little else to do, the group reminisces about various situations that they've been in. After a period of time, power is restored.

## Cast

* Soleil Moon Frye as Punky Brewster
* Cherie Johnson as Cherie Johnson
* George Gaynes as Henry Warnimont
* Susie Garrett as Betty Johnson
* Ami Foster as Margaux Kramer
* Sandy as Brandon "the Wonder Dog"
Concept Lab Prep Instructions

## Major Items to Prepare

1. Agar Cubes: 2+ days prior
2. Pasteur's Experiment x 3: 2 weeks prior
3. Copy Safety Agreements
4. Extra copies of Concept lab for students who have not yet bought the manual

## Agar Cubes

### **About**

These will be used in Exercise 1, as an example of surface to volume ratio. In this exercise, the students will soak the 2 sized cubes in an acid (25% vinegar) which will make the purple indicator turn clear from the outside-in. The higher the surface to volume ration, the faster the cube will turn clear. This is an exercise to demonstrate many things in biology having to do with surface area, particularly the small size of a cell.

### **Instructions**

\*Prepare 6 large (~2cm) cubes and 6 small (~1cm) cubes for each class section + some extras for good measure

**Materials**:

* Bromeresol Purple .4% (0.4g powder + 99.6mL H2O)
* 10g agar
* 450 mL DI H2O
* 50mL .01M NaOH
* 5 mL Bromocresol Purple

**Methods**

1. In 500mL media bottle, add 10g of agar powder
2. Add 395mL H2O
3. To make cubes, melt the agar,
4. Add 50mL .01M NaOH, and 5 mL Bromocresol Purple
5. Stir for hours or overnight to fully dissolve the indicator
6. Autoclave and store
7. To make cubes, put the bottle in a warm bath to melt the agar
8. Once melted, mix gently and pour into square container
9. Let sit or refrigerate until solid
10. Remove from container
11. Cut 2 cm and 1 cm cubes (I haven't found a great way to do this yet so do the best you can).

#### **Location of Materials**

* Bromocresol Purple: Chemical Cupboard C9
* Agar: Media cupboard C10
* Cube Tray: Drawer next to Bench sink?
* NaOH: Base cupboard in Room 317

**Instructions Improvement Notes**

Add an approximation of how much this recipe will make

## Pasteur's Experiment

### **About**

This is an amazing experiment that covers the scientific method. This is for Exercise 2.

### **Instructions**

Prepare  ***3 SETS*** of the experiment. There will be 3 stations in each lab for this one single exercise.

**Materials and Methods**

Prepare 18 x 500mL flasks with 200mL nutrient broth in each. You only need 15 but I always make a couple extra just in case of contamination. Follow the instructions on the bottle but...

**Nutrient Broth**

(These are instructions for 1 L of broth. I usually fix up 3600mL for about 18 flasks)

1. Suspend 8 g of nutrient broth powder in 1L of DI water.
2. Mix thoroughly on a hot plate
3. Boil for 1 minute to dissolve the powder
4. Aloquot 200mL into individual 500 mL flasks
5. Autoclave all flasks with stoppers on liquid for 15 minutes (121 degrees)
6. All stoppers and tubes that will be used must be autoclaved too
7. Immediately put appropriate stoppers and tubes on the flasks after autoclaving

**Flasks**

* A: Inoculate 3 open flasks with yucky swabs from yucky places and incubate
* B: Pour 3 into new, un‐autoclaved flasks and close
* C: Keep 3 Sealed completely with #7 stoppers, do not open after leaving the autoclave
* D: Affix 3 with straight glass tubes before autoclaving
* E: Affix 3 with hook tubes before autoclaving

### **Location of Materials**

* Nutrient agar: Media Cupboard C10

Original Notes\*
Tree Frogs

## About

Tree frogs are very cute and charming little frogs. They are pretty simple to care for and amazing to observe! Overall they are cool little animals that can't be handled but are a great joy to watch hop about.

 Quick Facts

 **Class** | Amphibia

* **Diet:** Insectivore
* **Lifespan:** ~5 years
* **Temp:** 45-60 Fahrenheit ideal, tolerant of a wide range
* **Habitat:** Lives near freshwater sources
* **Humidity:** ~80% is ideal, tolerant

## Feeding

**Adult Frogs:**

1. Locate fruit fly container.
2. Open frog house, after checking to make sure no frogs are near the top.
3. Tap fruit fly container to get flies to go to the bottom, then open it and let some fly in.
4. Close fruit fly container (after tapping it again to get all the flies to the bottom)
5. Done!

**Tadpoles:**

1. Remove boiled dandelions from fridge.
2. Place one in the tadpole tank.
3. Done!

## Care

**Adult frogs:**

* Spray enclosure with water daily, or as often as possible. Moisten the soil.
* Check water, make sure it is clean. If not clean, siphon water out and replace. DI water only; frogs are very sensitive to additives.
* Feed frogs as often as possible. Every other day is ideal.
* Watch the frogs! They are cute and entertaining when eating.

**Tadpoles:**

* Check water and make sure its clean. If not clean, siphon the gunk out of the bottom and put in new water.
* Feed when the old dandelion is gone and eaten.
* Check for legs. Move immediately if frog has grown both pairs of legs.

## Links & Resources

* Basic care info
* Species info
* Care sheet
* Wikihow article
Bio 10 Meiosis Lab Set-Up

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 Instructor Notes

**Meiosis!!!**

There are no really special instructions for this lab but I have included on your bench, a Meiosis quiz and the large meiosis worksheets. On the back bench, I have
computer animations that may help students visualize the process.

After class, please have students return the lab to how it was originally!

Thank you,

Danielle

## Documents

Meiosis Quiz

Meiosis Worksheet 11 x 17"
Simple Stain and Smear Prep

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**I. Smear Preparation and Simple Staining**
**Materials per table**

*24- to 48-hour nutrient broth* of Bacillus subtilis and/or E. coli,
*Staphylococcus aureus. If no Bacillus, no problem.*
* clean microscope slides
* bibulous paper
* various stains
* lens paper and lens cleaner

**\*\*grow B.s. at room temperature for 48 hours. Grows nicely on BHI or NB**
Concept Lab Instructor Notes

**Welcome to a new semester!**

**Day 1 obligations:**

1. Copies of Lab 1 are located on the instructor table. If students

 take a copy, please have them give you their copy when they buy the lab
 manual.

1. Be sure to go over the **Student Laboratory Rules Agreement** with your students. Have students sign the signature sheet and return it to me. Students under 18 years of age need to have a parent’s signature on the paper copy of the Lab Safety Agreement and it return it to me for our records.

1. I have also provided an introduction to the students with all of

 my information and services. I would be happy to come into the class to say hello and go over all of the information. Let me know! Spring and Fall semesters only: Please announce **Review Day** with Danielle, These are every Friday before Lab Exams from 10a to 4p.
**A few lab reminders:**

1. Backpacks will be placed in the cubbies each lab session
2. No food or drink in the labs
3. Please point out the location of the fire extinguisher, eye wash, shower, and evacuation area.
4. Please remind students to not eat or drink chemicals (unless directed by their instructors)
5. For Concept 1- each group of 4 students will use 1 large cube and 1 small cube (I have provided a total of 6 large and 6 small for the class).
6. For Exercise 1A: 2 stations. For Exercise 2 A&B: 3 stations

**Materials can be Found!**

If I am not here to set out all materials, here is where you can locate your materials:

1. 6 large and 6 small cubes are located in the prep fridge in their own container. Put 3 of each at the two student stations. There are extras in the fridge if needed.
2. Extra 25% vinegar is on your bench

**After you are finished:**

1. When the lab is finished, please dispose of the Waste cubes and vinegar in the waste bowls on your bench.
2. Please have students wipe down the benches, push in chairs, straighten exercise stations, and return everything how it was found.

Thank you!!!
Koch’s Postulates

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#### **Prep (Lab 1)**

Purchase 1 quart organic milk, usually Straus, Maria asked me to avoid Clover b/c it had failed in cheesemaking, Yogourmet starter.

1. Autoclave micro-centrifuge tubes, yellow for yogurt, white for milk. Will need one per pair of each in lab 1. If 48 students in two sections, then need ~25 or so of each.
2. Sterilize agar -- pour plates. Beth used a Tryptic Soy Agar Plus -- the plus meaning a recipe booster she uses. I have used for several years now a mixture of Tryptic Soy Agar mixed with Lactose Broth, following amounts called for on bottles -- i.e., if 20g/L for TSA & 5 g/L for LB, I use 20 + 5 for a liter. Make double the plates as they are used in Lab 1 (1 plate per pair) and Lab 3 (1 plate per pair).
3. Autoclave about 25 empty 12x125 test tubes with caps. A dozen extra minimum is good in case students find different types of colonies and they want to inoculate both.
4. Autoclave several beakers with stirring rods, covered with foil. Enough for a couple to be Bio10 yogurt (if Danielle wants it). A 1 liter, 2 125 ml, 1 500 ml, 1 250 ml (no
5. Heat milk in 1 liter

#### **Lab I (per pair)**

* 1 microtube holders
* 1 microtubes of sterile milk
* 1 microtubes (sterile) of yogurt
* 1 TSA+ lactose plate
* 1 test tube of sterile water
* 2 pH strips 2-7pH………….put guides out on back counter.
* loops

#### **Lab II**

* 1 tt with sterile milk (5mL each)
* loops

#### **Lab III**

* The 5mL tt of inoculated and incubated milk
* TSA+ plate
* loops
* pH strips 2-7pH……………put guides out on back counter.
* Wood stirrers
* tt of water for wet mounts

#### **Lab IV**

* Incubated plates
* staining supplies
* Sterile H2O
Punky's Millions

**"Punky's Millions"** was the second segment on the first episode of the second season of the cartoon spinoff series *It's Punky Brewster*.

## Synopsis

On a game show that Henry and Punky are competing on, they are informed that they have a chance to win millions of dollars if they can spend one million dollars in one week. However, because they can only spend a certain amount on each item, they are forced to come up with innovative solutions to the problems they face. For example, Glomer must find a way to make a racehorse lose, while Punky must find a variety of ways to assist a Catholic priest's orphanage. When they made their final appearance on the game show, Punky and the kids were required to demonstrate that they had spent the full one million dollars that they were required to spend. However, a mishap caused by Allen, in which a few cents in change fell out of his pocket, caused Punky and Henry to lose the game and be disqualified. Fortunately, the Catholic priest went to see Henry and Punky to let them know that he won to top prize on the lottery ticket Punky had purchased for him, and he decided to offer a sum of money to assist in the effort to save the orphanage. Punky and Henry come to the conclusion that they are winners after all.

## Trivia

* This segment is based on the 1902 George McCutcheon novel *Brewster's Millions*, which was adapted into a play in 1905 and a film in 1985.
* This is the second of two episodes in the series which involved gameplay. The other was "Winning isn't Everything".
Micro 5 Antibiotic Sensitivity

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### **Period 1**

* 4 broths of each: *Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus epidemidis, Enterococcus fecalis*
	+ All of these cultures should be between 48-72 hours.

* Sterile dry swabs, 3 swabs/per bench = 3x6=18/class
* little beakers for antiB to be exchanged Labeled "A, B, C, D, E" = 5 (1 for every 2 antiB's

* Antibiotic Sensitivity Discs and small single dispensers
	+ Amoxicillin
	+ Ciprofloxacin
	+ Erythromycin
	+ Gentamicin
	+ Penicillin
	+ Polymyxin B
	+ Streptomycin
	+ Tetracycline
	+ Vancomycin

I know I use one more.  Perhaps Ampicillin.  If we have Bacitracin, I would choose that one.

I am flexible and can use what we have available.

* Nutrient agar plates 6 per bench / group = 36 (6 groups)-42 (7 groups)
* forceps = 2/ bench

### **Period 2**

* Plates from period 1 (24/48 hrs in incubator?)

## Original (Lab Manual ) p. 103

### Lab 1

#### Per bench

* 1 broth *Staphylococcus aureus*
* 1 broth *Escherichia coli*

#### Per pair of Students

* 2 NA plates
* 2 sterile swabs dry

#### For the class antibiotic dispensing

* Antibiotic Disc Dispenser
* Alcohol jar
* forceps
* Antibiotics
	+ Ampicillin
	+ Chloramphenicol
	+ Erythromycin
	+ Gentamicin
	+ Penicillin G
	+ Polymyxin B
	+ Streptomycin
	+ Tetracycline

### Lab 2

* Incubated Plates
* rulers
Bench Top Sanitation

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**Student pairs**

* 4 NA plates (can use their plates they poured)
* 4 sterile swabs
* 2 Tiles

**Other**

* *E.c*. and *S.a.* tubes (for inoculation)
* Inoculation Swabs

**Solutions**

* Vesphine
* 10% bleach (2 squirt bottles)
* 70% ethanol (2 squirt bottles)
* Clorox wipes or Lysol wipes
* Other sprays....