pipetting toolS

EXPERIMENTAL objectiveS

To demonstrate the validity of Transcriptic’s Autoprotocol-implemented High Throughput Pipetting Quick Instruction.

methodology

Orange G electrophoresis dye (CHEM IMPEX) solutions of 33, 67 and 100 µg/ml were prepared in distilled water.  Thirty microliters of dye were transferred using the High Throughput Pipetting Quick Instruction from 96-well source plates to 384-well analysis plates and underwent absorbance measurement (475 nm) as an assessment of pipetting accuracy. 

Figure 1 Schematic of plate source/destination co-ordinates. Distilled water dye solutions were transferred from 96-well source plates using the High Throughput Pipetting Quick Instruction to quadrants 0, 1, 2 and 3 (denoted e.g. as “Q0” etc.) of 384-well analysis plates. Absorbance measurements at 475 nm were used to assess pipetting uniformity across wells.

Figure 1 Schematic of plate source/destination co-ordinates. Distilled water dye solutions were transferred from 96-well source plates using the High Throughput Pipetting Quick Instruction to quadrants 0, 1, 2 and 3 (denoted e.g. as “Q0” etc.) of 384-well analysis plates. Absorbance measurements at 475 nm were used to assess pipetting uniformity across wells.

results

Replicate transfers (n=96) performed using the High Throughput Pipetting Quick Instruction achieved minimal standard deviations for all 4 solutions (Figure 2). A linear relationship between absorbance and Orange G volume is observed with increasing dye concentration. 

Figure 2 (A)  Statistical analysis of absorbance readings (475 nm) for the 96 wells comprising each quadrant. S.D.: standard deviation. C.V.: coefficient of variance. (B) Heat map representation of absorbance across 384 wells. Distilled water was transferred to Q0, 33 µg/ ml, 67 µg/ml and 100 µg/ml Orange G solution to quadrants Q1, Q2 and Q3 respectively. (C) Box-plot representation of all 384 data points Whiskers: maximum/minimum values. Top boundary: 75th percentile. Bottom boundary: 25th percentile. Box divider: median value.

Figure 2 (A)  Statistical analysis of absorbance readings (475 nm) for the 96 wells comprising each quadrant. S.D.: standard deviation. C.V.: coefficient of variance. (B) Heat map representation of absorbance across 384 wells. Distilled water was transferred to Q0, 33 µg/ ml, 67 µg/ml and 100 µg/ml Orange G solution to quadrants Q1, Q2 and Q3 respectively. (C) Box-plot representation of all 384 data points Whiskers: maximum/minimum values. Top boundary: 75th percentile. Bottom boundary: 25th percentile. Box divider: median value.


VALIDATION DATE:  September 2015