Benefits

 

• High accurate and precise serial dilution can be implemented on EzMate 401 Automated Pipetting System with 8-channel Automated Pipetting Module

• Ideal for regular liquid handling task with walkaway convenience

 

Introduction

 

The highly accurate and precise EzMate Automated Pipetting System is specialized in handling low-volume pipetting tasks. The highly uniform 8-channel Automated Pipetting Module (APM) makes it an ideal tool for fast, accurate pipetting. In this application note, we demonstrated a consistent serial dilution operation using EzMate 401 Automated Pipetting System.

 

Material

 

Equipments:

 

• EzMate 401 with 8-channel 50 μl Automated Pipetting Module (APM50-8)
• Roche LightCycler®  480 Real-Time PCR System

 

Reagent:

 

• Finnzymes DyNAmo SYBR Green qPCR Kit, # F-415-L

 

Template:

 

• 101 bp synthetic DNA with following sequence, 1X working Conc. 1 pM:
• AAC TTG GCT TTA ATG GAC CTC CAA TTT TGA GTG TGC ACA AGC TAT AGA ACA CCA
• CGT AAG ACA TAA AAC GGC CAC ATA TGG TGC CAT GTA AGG ATG AAT GT

 

Primers:

 

• Forward :AAC TTG GCT TTA ATG GAC CTC CA, working Conc. 300 nM
• Reverse:  ACA TTC ATC CTT ACA TGG CAC CA, working Conc. 300 nM

 

Consumables:

 

• Roche LightCycler®  480 Multiwell Plates 384, # 047729749001
• Axygen®  50 ul Robotic Tip, w/o filter, Non-Sterile, # FX-50-R

 

Methods

 

1. Prepare 3 sets of 4 fold serial dilution separately, starting from 1 pM DNA. (figure 1.)     

 

                                        Figure 1

 

2. Prepare the pre-mix solution (listed in the table 1 ). Use EzMate 401 with 8-channel 50 μl Automated Pipetting Module (APM50-8) to dispense 18 μl/well of the premix solution into each well in a clean 384-well qPCR plate.    

 

 

                                      Table 1. Pre-mix solution

 

3. Use EzMate 401 with APM50-8 to dispense 2 μl of the diluted DNA into each well of the 384-well plate (blue frames in figure 2 below for dilution series 1, red frames for dilution series 2 and green frames for dilution series3), triplicate 8-well for each concentration.

 

                                                            Figure 2

 

4. After dispensing, seal the plate, gently tap its side and then centrifuge it to remove any bubbles in the well. Run the qPCR experiment under the following cycling conditions: (table 2)

 

                                              Table 2. Cycling conditions

 

Result

 

1. qPCR results

 

Figure 3. Amplification curve of Roche LightCycler®  480

 

Figure 4. Standard Curve for all dilutions

 

 

2. The standard deviations for the Cps for each concentration replicates are less than 0.5.

3. The R2 values for all the standard curves are greater than 0.98.

4. The PCR efficiencies are all within 100% ± 10%.

 

Conclusion

 

The high accuracy and precision of the EzMate automated pipetting system in μl-level liquid pipetting combined with the uniform 8-channel Automated Pipetting Module make it an ideal tool for fast and consistent pipetting task. The application-oriented EzStarter control software is specially designed to meet modern day molecular biology laboratories’ need.

 

*Trademarks: Roche LightCycler® (Roche Group); DyNAmo™ (Finnzyme Oy); Axygen® (Corning Inc.)

 

DOWNLOAD PPLICATION NOTE

 

 

Sample Preparation Related Applications

 

• Sample Normalizations Performed by EzMate 401 Automated Pipetting System

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Learn More About EzMate Automated Pipetting System

 

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