# Standard Operating Procedure: General Laboratory Safety
## ENTM 201L - Molecular Biology Laboratory
### University of California, Riverside

**Document ID**: SOP-00
**Version**: 1.0
**Effective Date**: Fall 2025
**Author**: Luciano Cosme

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## 🎯 Purpose & Scope

This SOP establishes fundamental safety practices for all personnel working in the ENTM 201L molecular biology teaching laboratory. These guidelines apply to **everyone** - students, instructors, and visitors - during all laboratory sessions involving DNA extraction, PCR, gel electrophoresis, and molecular diagnostics.

**Remember**: Safety isn't about following rules - it's about going home healthy every day.

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## 🛡️ OUR SAFETY-FIRST APPROACH

### Pre-Aliquoting System for Maximum Protection

**EVERYTHING IS PRE-PREPARED FOR YOU!**

To minimize risk and maximize learning, your instructor:
- ✅ Pre-aliquots all chemicals into labeled tubes
- ✅ Prepares exact volumes needed for each experiment
- ✅ Pre-pours all gels with GelRed stain
- ✅ Dilutes all buffers to working concentrations
- ✅ Handles all hazardous waste disposal

**This means you can focus on learning techniques without chemical exposure risks!**

Your job is simple:
1. Follow the protocol cards
2. Use what's provided
3. Ask if unsure
4. Never open stock bottles

---

## ⚠️ Hazard Overview - MINIMIZED BY DESIGN

### Our Safety Philosophy:
**The instructor pre-aliquots ALL chemicals and prepares ALL hazardous materials. Students work only with pre-prepared, labeled materials in safe quantities.**

### What We're Working With:
- **Biological**: Preserved mosquito specimens (potential pathogens, though minimal risk)
- **Chemical**: Pre-aliquoted DNA extraction buffers, diluted ethanol, pre-poured gels with stain
- **Physical**: Centrifuges (12,000 rpm), UV light, electrical equipment
- **Sharp Objects**: Forceps and micropestles

### What Students NEVER Handle:
- ❌ Stock chemicals or concentrated buffers
- ❌ Pure GelRed or other DNA stains
- ❌ Undiluted acids or bases
- ❌ Hazardous waste disposal

---

## 👔 Before You Enter: Dress Code

### ✅ Required Attire
- **Closed-toe shoes** (NO sandals, Crocs, or open-heel shoes)
- **Long pants** (NO shorts, skirts above ankle, or leggings as sole layer)
- **Shirts with sleeves** (NO tank tops or exposed midriffs)
- **Hair tied back** if below shoulder length

### ❌ Remove Before Lab
- Dangling jewelry (earrings, necklaces, bracelets)
- Loose scarves or ties
- Bulky rings that interfere with gloves
- Headphones/earbuds

### 💡 Pro Tips:
- Wear old clothes - stains happen!
- Bring a hair tie even if you "think" your hair is short enough
- Remove nail polish if chipped (flakes contaminate samples)

---

## 🥽 Personal Protective Equipment (PPE)

### The Essential Four:

1. **Lab Coat** 👔
   - Button/zip completely
   - Sleeves rolled down
   - Remove before leaving lab
   - Never wear in eating areas

2. **Safety Glasses** 👓
   - ANSI Z87.1 rated
   - Over prescription glasses
   - Side shields required
   - Clean with lens paper only

3. **Nitrile Gloves** 🧤
   - Change when contaminated
   - Change between different samples
   - Never touch face with gloves
   - Size matters - too big = poor dexterity


### The Glove Rules That Save Experiments:

**ONE GLOVE RULE**: Use one glove for "dirty" work, keep one hand clean for phones

**NEVER touch with gloves on:**
- Your phone 📱
- Computer keyboards ⌨️
- Door handles 🚪
- Light switches 💡
- Your face 😊
- Your water bottle 💧

**ALWAYS change gloves after:**
- Handling each mosquito specimen
- Spilling anything
- Touching non-sterile surfaces
- Sneezing/coughing into hands
- Every 2 hours (they degrade)

---

## 🧫 Working with Mosquito Specimens

### Safe Handling Procedures:

1. **Forceps Technique** 🔧
   - Hold like a pencil, not a hammer
   - Never point toward faces
   - Place in designated holder when not in use

2. **Pestle Grinding** 💪
   - Use inside tube only
   - Twist, don't pound
   - One pestle per sample (no sharing!)
   - Dispose in sharps container if glass

3. **Preserved Specimens** 🦟
   - **Frozen (-80°C)**: Keep on ice, work quickly
   - **Ethanol (95%)**: Allow to evaporate before grinding
   - **Silica gel**: Remove beads or mosquito with forceps

### Cross-Contamination Prevention:
```
Sample 1 → Clean forceps → Sample 2 → Clean forceps → Sample 3
         ↑                           ↑
    70% ethanol                70% ethanol
```

---

## 🧪 Chemical Safety - PRE-ALIQUOTED FOR YOUR PROTECTION

### How We Keep You Safe:
**ALL chemicals are pre-aliquoted by the instructor into labeled tubes with exact amounts needed. You will NEVER handle stock bottles or measure hazardous chemicals.**

### What You Actually Handle (Pre-prepared):

| What Students Receive | Original Hazard | Your Risk Level |
|----------------------|-----------------|-----------------|
| **Lysis buffer** (300 μL tubes) | Corrosive (concentrated) | Minimal (diluted, small volume) |
| **Ethanol** (pre-measured) | Flammable | Low (small aliquots) |
| **Gels** (pre-poured with GelRed) | Mutagen | None (already in gel) |
| **TAE buffer** (ready to use) | Irritant (concentrated) | None (diluted 1X) |
| **Proteinase K** (10 μL aliquots) | Sensitizer | Minimal (tiny amount) |

### The Spill Response: **S.P.I.L.L.**
- **S**top and alert others
- **P**rotect yourself (PPE)
- **I**solate the area
- **L**imit spread with absorbent
- **L**eave if beyond your training

**Small spills (<100 mL)**: Use spill kit
**Large spills (>100 mL)**: Call EHS (951) 827-5528

---

## ⚡ Equipment Safety

### Centrifuge Operation 🌀
- **ALWAYS balance tubes** (opposite positions, equal weight)
- Check rotor is secure
- Close lid completely
- Stay nearby during first minute
- If wobbling: STOP immediately
- Never open while spinning

### Thermocycler/Heat Blocks 🌡️
- Heated lid = 105°C (will burn!)
- Use tube lifter for hot samples
- Never leave tubes uncapped at high temps
- Check program before starting

### Gel Electrophoresis ⚡
- Always run "away from you" (black to red)
- Never touch buffer while running
- Use one hand rule when connecting
- Cover tank while running
- Max voltage: 100V for our gels

### UV Transilluminator 🟦
- Face shield required (not just safety glasses)
- Minimize exposure time
- Close door/shield
- Never look directly at UV
- Photo quickly, analyze on computer

---

## 🗑️ Waste Disposal - SIMPLIFIED FOR SAFETY

### Student Responsibilities (Simple!):

1. **Biological Waste** 🧬
   - Used pipette tips → Red biohazard bag on your bench
   - Mosquito remains → Red biohazard bag
   - That's it!

2. **Regular Trash** 🗑️
   - Clean paper towels → Regular trash
   - Empty tubes (if dry) → Regular trash
   - Clean gloves → Regular trash

### Instructor Handles ALL:
- ✅ Chemical waste disposal
- ✅ Hazardous buffer disposal
- ✅ Gel disposal with GelRed
- ✅ Sharps container management
- ✅ Waste pickup requests
- ✅ Ethanol waste

**Students NEVER:**
- ❌ Pour anything down the drain
- ❌ Handle chemical waste containers
- ❌ Dispose of sharps
- ❌ Clean up chemical spills alone

**When in doubt, ASK - don't guess!**

---

## 🚫 Prohibited Activities

### The Absolute No's:
- ❌ **No eating, drinking, or gum** (not even water!)
- ❌ **No applying cosmetics** (includes lip balm, hand lotion)
- ❌ **No mouth pipetting** (use mechanical pipettes only)
- ❌ **No horseplay or pranks** (equipment is expensive and dangerous)
- ❌ **No unauthorized experiments** (stick to protocols)
- ❌ **No working alone** (buddy system required)
- ❌ **No shorts or sandals** (even "just for a minute")

### The "Ask First" List:
- 🤔 Taking photos for reports
- 🤔 Adjusting equipment settings
- 🤔 Using different reagents
- 🤔 Staying late to finish

---

## 🧹 Workspace Management

### Start of Lab: **The 3-Check System**
1. ✓ **Check equipment** - Everything working?
2. ✓ **Check supplies** - Everything available?
3. ✓ **Check surface** - Clean with 70% ethanol

### During Lab: **The Clean-As-You-Go Rule**
- Wipe spills immediately
- Return shared items promptly
- Keep only current materials on bench
- Label everything immediately

### End of Lab: **The 5-Point Shutdown**
1. ⬜ Turn off all equipment
2. ⬜ Properly dispose all waste
3. ⬜ Clean bench with 70% ethanol
4. ⬜ Wash and store reusable items
5. ⬜ Remove PPE and wash hands

---

## 🤝 Lab Etiquette

### Be a Good Lab Citizen:
- 📢 Communicate spills/breaks immediately
- 🏷️ Label with: Name, Date, Contents
- ⏰ Be on time (safety briefings are mandatory)
- 📱 Phones on silent, no calls in lab
- 🎵 No music without instructor permission
- 🤧 Stay home if sick

### Shared Equipment Rules:
- Clean before and after use
- Report problems immediately
- Don't hog equipment
- Ask before adjusting settings

---

## 🚨 Emergency Procedures

### Know These Locations:
- 🚿 **Safety shower**: _____________
- 👁️ **Eyewash station**: _____________
- 🧯 **Fire extinguisher**: _____________
- 🏥 **First aid kit**: _____________
- 🚪 **Emergency exits**: 2 minimum
- 📞 **Emergency phone**: _____________

### Emergency Contacts:
| Emergency Type | Contact | Number |
|---------------|---------|--------|
| **Life-threatening** | 911 | 911 |
| **UCR Police** | UCPD | (951) 827-5222 |
| **EHS** | Environmental Health & Safety | (951) 827-5528 |
| **Poison Control** | California Poison Control | 1-800-222-1222 |
| **Instructor** | Cosme | ___________ |


### Specific Emergency Responses:

#### Fire 🔥
- **R.A.C.E.**
  - **R**escue anyone in danger
  - **A**larm - pull station
  - **C**ontain - close doors
  - **E**vacuate/Extinguish (if small)

#### Chemical Exposure 💧
- **Skin**: Flush 15 minutes minimum
- **Eyes**: Eyewash 15 minutes, get medical attention
- **Inhalation**: Move to fresh air
- **Ingestion**: Call Poison Control, never induce vomiting

#### Injury 🩹
- **Minor cuts**: Clean, bandage, report
- **Major cuts**: Apply pressure, call 911
- **Burns**: Cool water (not ice), seek medical attention
- **Needlestick**: Wash, report immediately

---

## 📝 Training & Documentation

### Required Training:
- [ ] This SOP (read and sign)
- [ ] Lab-specific safety orientation
- [ ] PPE training via LHAT
- [ ] Waste disposal training
- [ ] Emergency response walkthrough

### Documentation:
- Sign-in sheet each lab session
- Incident reports within 24 hours
- Near-miss reports encouraged
- Training records maintained 3 years

---

## 🎓 Special Considerations for Teaching Lab

### Common Student Mistakes to Prevent:
1. **Forgetting to balance centrifuge** → Post reminder signs
2. **Contaminating stock solutions** → Use aliquots only
3. **Mixing up samples** → Labeling practice first day
4. **Pipetting errors** → Practice session required
5. **Phone contamination** → Designated phone zones

### Creating a Safety Culture:
- Mistakes happen - report them!
- Questions are encouraged
- Safety suggestions welcome
- Peer reminders are helpful
- Excellence includes safety

---

## ✅ Quick Safety Checklist

**Before Lab:**
- [ ] Appropriate clothing
- [ ] Hair tied back
- [ ] Jewelry removed
- [ ] Know emergency exits

**Starting Work:**
- [ ] PPE on correctly
- [ ] Workspace cleaned
- [ ] Materials labeled
- [ ] Protocols ready

**During Work:**
- [ ] One glove rule
- [ ] Clean as you go
- [ ] Communicate issues
- [ ] Follow protocols

**Leaving Lab:**
- [ ] Equipment off
- [ ] Waste disposed
- [ ] Bench cleaned
- [ ] Hands washed

---

## 📋 Agreement & Acknowledgment

By signing below, I acknowledge that I have:
- Read and understood this SOP
- Had opportunity to ask questions
- Agree to follow all procedures
- Will report unsafe conditions
- Understand consequences of non-compliance

**Name**: _________________________________

**Signature**: _____________________________

**Date**: __________________________________

**Instructor Verification**: __________________

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## 📚 References

1. UCR Environmental Health & Safety Laboratory Safety Manual
2. CDC Biosafety in Microbiological and Biomedical Laboratories (BMBL) 6th Edition
3. ASM Guidelines for Biosafety in Teaching Laboratories
4. Zymo Research Quick-DNA Tissue/Insect Kit Safety Data Sheet
5. BioDynami HMW Magnetic Bead Kit Safety Data Sheet
6. California Code of Regulations, Title 8, Section 5191

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## 📝 Revision History

| Version | Date | Changes | Author |
|---------|------|---------|--------|
| 1.0 | Fall 2025 | Initial version | L. Cosme |

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*"Safety is not a gadget but a state of mind." - Eleanor Everet*

**Questions? Concerns? Suggestions?**
Contact: Luciano Cosme
Email: luciano.cosme@ucr.edu
Office: ENTM 325