improving the molecular biology workflow

Streamlining the Molecular Biology Workflow

Labnet International Presents

May 2, 2023 Labnet International - Molecular Biology Workflow 2

The Molecular Biology Workflow

The Molecular Biology Workflow Involves 4 Key Steps:

1. Sample Prep2. PCR Prep3. PCR4. Validation

Overview


Liquid Handling is needed throughout the Molecular Biology workflow, from Sample Prep to PCR.

Reliable and accurate liquid handling is critical.

Liquid Handling


• Personnel → Good pipetting technique comes from practice

• Facility → Environmental conditions• Apparatus → Right instrument• Procedures → Regular Maintenance • Material and reagents → Performance

checks (calibration)

Good Pipetting Procedures

Liquid Handling


SET THE VOLUME

• Using a push button or a knob

• Always possible with one or two hands and with gloves

AVOID PARALLAX ERROR

• Horizontal position• Volume lock• Approach a volume from

a higher value

Setting the Volume

Liquid Handling


ASPIRATE AND DISPENSE BACK BEFORE USE!

Why?• Greater uniformity• Great accuracy and

precision

When?• After changing the tip• After increasing the

volume

Pre-rinsing – Wet Tip

Liquid Handling


1. Results obtained by untrained technician (red curve)

2. Results obtained by a trained technician (green curve)

• T: Target – Set Volume• R: Volume range –

permitted by the manufacturer specification

Personnel - User Mistakes Influence Results

Liquid Handling


1. Sample Prep


Prism™ R Refrigerated Microcentrifuge

• Powerful Refrigeration attains 4°C in just 8 minutes

• Compact footprint - only 25.4 cm wide

• Aluminum rotor• 24 x 1.5/2.0 ml tubes

or PCR strips• Easy Access Design• Optional Strip Spin

Adapter

Sample Prep


Axygen Mini-Prep Multisource Genomic DNA Kit

• Mammalian Tissue• Plant• Cells• Fungi

Sample Prep

Transfer Miniprep column to a 2 ml microfuge tube and centrifuge for 1 minute at 12,000 x g.


Prism™ R Easy Tube Access

Samples are easy to insert and remove from the rotor.

Sample Prep


Prism™ R Rapid Acceleration and Deceleration

Maximum G-Force: 17,135 x gMaximum RPM: 13,500 rpm

Sample Prep


Prism™ R Reduces Rotor Handling

24 x 1.5/2.0ml aluminum rotor

Sample Prep

24 x 1.5/2.0ml aluminum rotor

Additional adapters are available for spinning tubes smaller than 1.5ml


2. PCR Prep


IMPROVES REPRODUCIBILITY AND STREAMLINES WORKFLOW FEATURES AND BENEFITS

• Quickly spins down droplets and condensation

• Max RPM = 2,500• Max G = 500 x g• Use before and after PCR

to improve yield and reduce noise

• Accepts skirted, non-skirted and all standard PCR plates

• Less than ¼ the size of standard plate centrifuges

MPS 1000 Mini Plate Spinner

PCR Prep


LESS THAN ¼ THE SIZE OF STANDARD PLATE CENTRIFUGES

MPS 1000 Mini Plate Spinner – Size Advantage

PCR Prep


Application: MPS 1000 Mini Plate Spinner Centrifuge for Real Time PCR

Studies show that using the MPS 1000 before PCR increases PCR yield and reduces “noise” in the dilution series.

PCR Prep


MPS 1000 Mini Plate Spinner – How it Works

1. Insert the Plates2. Spin for 20 seconds3. Remove the Plates

PCR Prep


Plate Adapter for MPS 1000 Mini Plate Spinner

Supports:• 8 x 0.2ml PCR Strips• 12 x 0.2ml PCR Strips• 0.2ml PCR Tubes• 4 x 6 half plates

PCR Prep


3. PCR


AccuSeal™ Semi-Automated Plate Sealer

The new Labnet AccuSeal is a semi-automated heat sealer designed to seal a wide variety of microplates using different types of heat seal films.

PCR


• Consistent sealing across all wells• Consistent results plate to plate• Less chance of leakage or evaporation• Better integrity at low temperatures (-80°C

freezer storage)• A variety of types of seals for flexibility with

applications• Easy to operate – set the parameters

correctly and consistent results are virtually guaranteed

Advantages of Thermal Sealing

PCR


• PCR Thermal Cycler• 96 well format• Fast run times• “Better than

Gradient”• Upgrade of

MultiGene Gradient• 3 year warranty

MultiGene™ OptiMax Thermal Cycler

PCR


MultiGene™ OptiMax – Custom Optimization – “Better than Gradient”

PCR

1. Can set all wells for 1 temp2. Gradient above sets 1st

column and last column and cycler determines values in between

1. Can set all wells for 1 temp2. Gradient above sets 1st block and

last block and cycler determines values in between

3. User can set values of choice for each block – “Better than Gradient”


AS PUBLISHED IN MARCH 2013 BIOTECHNIQUES

Streamlining the Genotyping Workflow with the MultiGene™ OptiMax

PCR

http://www.biotechniques.com/BiotechniquesJournal/2013/March/Streamlining-the-Genotyping-Workflow-with-the-MultiGene-OptiMax/biotechniques-341103.html

May 2, 2023 Labnet International - Molecular Biology Workflow

P GENE

• 94°C for 3 minutes

35 cycles of:• 94°C for 30

seconds• 51.7°C for 1

minute• 68°C for 1 minuteThen• 68°C for 2

minutes• 4°C hold

MultiGene™ OptiMax

PCR

D GENE






T GENE







Comparisons of results with Traditional Thermal Cycler and OptiMax

PCR

1- Ladder2- WT (wild type)3- P transgene + WT4- P transgene + WTControls were from previously genotyped animals displaying either a P transgene or a Wild Type (WT) band. (Not depicted here)

1- Ladder1 – F1/F1 (homozygous of T gene)3- Het (one copy of T gene + WT)4- F1/F1 (homozygous of T gene)Controls were from previously genotyped animals displaying either a WT or both WT gene bands. (Not depicted here)


Comparisons of results with Traditional Cycler and OptiMax

PCR

1- Ladder2- DR/DR (homozygous of D gene)3- Het (one copy of D gene + WT)4- HETControls were from previously genotyped animals displaying either a homozygous for D gene or both WT and D gene bands. (Not depicted here)


SIMPLIFIED WORKFLOW – IMPROVED THROUGHPUT

The MultiGene OptiMax thermal cycler may be programmed to operate between one and six different annealing temperatures of user choice across the block.• Improved throughput by

reducing time• Simplified workflow by

reducing steps

MultiGene™ OptiMax

PCR

Fig. 1 Primers designed to anneal to various genes (e.g Gene A, B, and C) typically have different annealing temperatures (T1,T2 and T3). To simplify workflow and increase throughput, the MultiGene OptiMax can perform up to six different reactions at the same time thus reducing the time from 9+ hours to three.


Better than Gradient cyclers are ideal for Genotyping labs:1. Zebra fish2. Arabidopsis3. Drosophila4. Transgenic Mice

Technical / Applications

PCR


4. Validation


IMPROVES SPEED AND REDUCES SETUP TIME

1. Saves time2. Run entire 96 well plate

for quick screening3. Multichannel pipette

compatible

Enduro™ 96 Horizontal Gel Box

Validation


SPEED AND EASE OF USE

1. Blue light LEDs – Powerful EPI blue LEDs enable the user to capture beautiful images of SYBR safe stained gels

2. Tablet interface creating small footprint – system arrives fully assembled, with software preinstalled.

3. 3 position filter slider – increased application flexibilty – orange filter comes standard (green, red and blue filters are also available.)

Enduro™ GDS Touch Gel Documentation System

Validation


SPEED AND EASE OF USE

1. 4 clicks to results2. Small footprint3. Improved blue light

sensitivity4. Will never need to touch

camera5. Improves workflow

Enduro™ GDS Touch Gel Documentation System

Validation


GEL IMAGED WITH BLUE LIGHT CONVERSION SCREEN

GEL IMAGED BLUE LED – ENDURO™ GDS TOUCH

Enduro™ GDS Touch Applications

Validation

Blue LED provides greater sensitivity as a Blue Light Conversion Screen SYBR® Safe


• DNA gels• Protein gels• Stains including but not limited to:

• Ethidium Bromide• Coomassie Blue• Silver Stain• SYBR® Gold• SYBR® Green• SYBR® Safe• GelStar• SYPRO® Ruby

Enduro™ GDS Touch Applications

Validation


1. Centrifugation – Reduces cool down time

2. Liquid Handling – Good practices and easy self calibration

3. Plate Spinner – More reproducible results

4. Plate Sealer – More reproducible results

5. Thermal Cycler – Reduced time and steps

6. Gel Electrophoresis – Reduced preparation time

7. Gel Imaging – Reduced run time and improved results

Streamlining Molecular Biology Workflow

Conclusion


Learn More About Streamlining the Molecular Biology Workflow. Visit us at http://labnetinternational.comTwitter: @labnetnews

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improving the molecular biology workflow

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