GPP™ — Good Pipetting Practice™

 Reducing the risks of user in liquid handling

 Improve the accuracy and reproducibility

Internal usage only

GPP™ — Good Pipetting Practice™

Internal usage only

Evaluation and Selection

Sample evaluation
 Sample number
 Sample quantity
 Sample type

Internal usage only

Evaluation and Selection

Manual pipette

96-well pipette

Electronic pipette Internal usage only

Electronic vs Manual Pipette

 Benefits of Electronic pipette

- Highly consistent pipetting
- High accuracy and precision
- Advanced functionality (multi-
dispensing, serial dilution, selectable
speeds, etc)
- Highly ergonomic

 However…
- Higher cost
- Required to program pipette

7 Internal usage only

Choosing the Right Tip

 Design
 Quality
 Fit

8 Internal usage only

Standard Tip Design

 Fine point, thin walled

 Clear
 Good sealing

9 Internal usage only

Specialty Tip Designs
Filter
Aerosol-resistant

Low-retention

Extended length

Wide orifice

Shaft protection (ShaftGard)

Gel loading (Gel-Well)

10 Internal usage only

Tip/Shaft Designs: LTS™ and Traditional

Small seal Large seal area

Friction-fit

Positive stop
In tip

LTS Cylindrical Traditional Conical

Dramatic reduction
in tip ejection force

11 Internal usage only

 Tip Quality

 For reliable results, tips should be:

- Clean to avoid contamination of sample by:
RNase, DNase
DNA
Pyrogen
ATP

- Inert to avoid addition of impurities from

polypropylene and plasticizer byproducts
Virgin polypropylene
No Oleamide, DiHEMDA

- Manufactured in an ISO 9001 Certified facility

Guaranteed of cleanliness

12 Internal usage only

Training

Pipetting Techniques
 Techniques to Reduce Errors
and Ensure Data Quality

13 Internal usage only

The Pipetting Cycle (Forward Pipetting)

14 Internal usage only

Setting the Micrometer

 Turn micrometer 1/3

revolution above desired
volume.
 Dial down to volume
setting.
 Approach each volume in
the same direction each
time.

Performing this technique can enhance performance by up to 0.5%

Correctly setting the micrometer improves accuracy up to 0.5%.

15 Internal usage only

Tip Immersion Angle

Incorrect Immersion Angle Correct Immersion Angle

Correct angle
Aspirating pipetting
with the enhances performance
pipette perpendicular by up
improves accuracy to0.5%,
up to 2.5%
ultramicro up to 2.5%.

16 Internal usage only

Tip Immersion Depth

0.1 -10 µL: 1-2 mm 10 -200 µL: 2-3 mm 200-2000 µL: 3-6 mm
Over 2000 µL: 6-10 mm

Recommended immersion depth improves accuracy up to 1%,

ultra-micro up to 5%.

17 Internal usage only

Aspiration Rate Effects

 Damage piston & seal

 Introduce aerosol & cross
contamination

Poor technique Good technique

Even without splashing, inconsistent aspiration can affect accuracy 1-5%.

18 Internal usage only

Pause Time for Macro-volume Pipettes

 Wait a minimum of 1 second with tip immersed

after aspiration
 Withdraw tip slowly and smoothly from liquid
 Important for large volume samples and viscous
samples

19 Internal usage only

Dispensing Techniques

 Three techniques:
1. Along side-wall (touch off)
2. Above liquid surface
3. Into liquid
 Key Considerations:
- Pause before blowout with
viscous liquids
- If dispensing in liquid, keep
piston at second stop until
tip is removed to prevent
unintentional aspiration

1 2 3

Ensuring complete dispensing can enhance performance up to 1%

20 Internal usage only

Pre-rinsing Pipette Tips

 Aspirate sample into tip, and

then dispense back into reservoir
or to waste
 Pre-rinsing provides identical
contact surfaces for all aliquots Press and release plunger

 Pre-rinse tip with same liquid that

two or three times

is being dispensed

2 pre-rinses provide up to 0.2% greater accuracy with aqueous liquids.

21 Internal usage only

 Consistency

 Use consistent
- Pipetting rhythm
- Pressure on plunger
- Speed and smoothness

 For best consistency

- Use an electronic pipette
Provides optimum consistency
Requires less user technique

22 Internal usage only

Avoid Hand-warming

To avoid hand warming the pipette, use a high-quality instrument and place it on a
stand when not in use.

To avoid hand warming the pipette, use a high-

q uality instrumentand place it on a stand when
not in use.

23 Internal usage only

Calibration and Routine Operation

Maintaining Peak Performance

 Quick Check/Verification
 Routine Maintenance
 Calibration

24 Internal usage only

Calibration

 Pipette performance evaluation

 Adjustment of a pipette’s calibration
setting, based on evaluation
 Fundamental compliance requirement
of external quality standards
(FDA, GMP/GLP, CLIA etc)
 Full value calibration includes
– As found performance data
– Preventive maintenance
– As returned performance data
– Documentation, labels
– Regularly scheduled intervals

25 Internal usage only

 Full-value Calibration Providers
Calibration
Calibration at Professional
Accredited Service Facilities
 Controlled environment
 Gravimetric balances: 6 and 7-place
 Highly trained and qualified staff
 Rigorous standards
 As found and as returned
documentation
 Sealing system parts replaced with
brand /model specific components
 ISO 17025 accredited
 Customer audits
 Routine preventive maintenance
included
Verification

 Two kinds of on site pipette

performance verification:
- Gravimetric Quick Check
- Leak Testing

VerificationVerification
results specify whetherspecify
results calibration is necessary.
whether calibration is necessary

27 Internal usage only

 Routine Maintenance

 Rainin’s 12 Point Pipette Inspection

 Basic Repair and Preventative Maintenance:
1. Shaft
2. Piston Assembly
3. Seal
4. O-Ring

 Cleaning and Decontamination

 Vacuum and Liquid Leak Testing

 Most Common Issues that Affect Pipette Performance

28 Internal usage only

 12 Point Inspection
1. Plunger Button
 Rainin 12 Point Visual Inspection 2. Plunger Rod
4. Volume Lock Top
1. Plunger button: Must be clean, window 3. Tip Ejector Button
display not cracked, & top piece turns freely. 5 & 6 Digital Volume
The top portion of button must spin freely Indicator/Micrometer
without adjusting the micrometer
2. Plunger Rod: Turns smoothly, not warped,
not bent, or rusted
3. Tip Ejector Button: Button moves up and
down freely. Does not stick or has sharp
edges. Must be clean
4. Volume Lock: Must not be cracked, split, or
loose. Must turn smoothly
5&6. Digital Volume Indicator/Micrometer:
Window is not cloudy, cracked, and debris on
micrometer dials. Check for vertical alignment
of dials. Numbers not faded or peeled

29 Internal usage only

 12 Point Inspection

 Rainin 12 Point Visual Inspection

7-9. Piston Assembly: This includes the
seal, o-ring, stroke spring, blowout spring,
large spring assembly holder, blowout
spring assembly holder, and seal assembly
holder.
 Piston free from rust or chemical splash
 Stroke spring not split
 O-ring (Black colored) not cracked 11. Shaft Coupling Nut
 Seal not cracked or split
 Seal Assembly holder not damaged
7-9 Piston Assembly:
 Large spring holder not damaged
10. Shaft: Clean, not pitted, scratched, worn, O-Ring, Seal, Seal
Retainer, &Stroke Spring,
clogged on the inside and outside
11. Shaft Coupling: Not cracked, threads
damaged, and holds shaft securely 10. Shaft
12. Tip Ejector Arm: Not rusted, not bent, no
sharp edges, and does not stick when in
use 12. Tip Ejector Arm

30 Internal usage only

 CLEANING AND DECON

1. Soft bristle toothbrush or small brush. Used

for scrubbing the pipette body.
2. Long stem cotton swabs. Used to clean hard
to reach places.
3
3. Use of Cole Palmer 8890 Ultra Sonic Bath
with warm water and Micro-90 solution.
 With Heating cycle and sonic vibration
 Great for cleaning shafts, tip ejector arms,
empty pipette body, shaft coupling nut.
4. IPA (Isopropyl Alcohol) used to wipe the
body, clean, and sterilize.
5&4
5. Acetone (TO BE USED SPARINGLY).
Great for removing glue residue.
6. DNA-OFF and Zap-Away: great for
removing DNA, RNA, RNase, DNase 1
2
7. 10% bleach and water. Rinse with Distilled
water and dry.

31 Internal usage only

 Leak Testers

1. Recommended for air displacement

pipettes in order to thoroughly test:
 Seal
 Shaft
 O-Ring
 Piston
 Tip

2. Liquid Leak Tests with:

- Di water
- Pancake Syrup
- Colored water (for multichannel) to
ensure even aspiration on all
channels.

32 Internal usage only

 Pipette Performance
 Factors That Could affect Pipette
Performance
1. Seals 60% 1) Not having pipette
serviced routinely. 2)Rusted piston
damages seal. 3) Piston too dry and
creates too much friction on seal.
2. Piston 25% 1) Rusted piston. 2)
Scratched Piston. 3) Bent Piston.
3. Shaft 10% 1) Scratched or pitted. 2)
Worn shaft. 3) clogged or dirty shaft.
4. Other 5% Can be found in the Pipette
handle or body, such as but not limited to:
1. Bent plunger rod
2. Micrometer over traveled (MOOP)
3. Calibration screw adjusted
4. Pipette dropped forcefully at correct
angle (accidentally).
5. Pipette autoclaved
5. The Volume Lock: Too loose, too tight, or
removed from pipette.

33 Internal usage only

34 Internal usage only