PLATEMASTER®: HIGH THROUGHPUT,MANUAL NGS LIBRARY PREPARATION
Krammes L., Roth T., Bauer E., Peter W., Schäfer M. | HLA-Laboratory, Stefan-Morsch-Stiftung, Birkenfeld, Germany
Binzer M., Dittmer J. | Gilson International BV, Germany

ABSTRACTPIPETTING TIME COMPARISONThe Stefan-Morsch-Stiftung (Foundation) screens the human leucocyte
anti­gen (HLA) gene complexes of potential stem cell donors to identify
potential organ donors for leukemia patients. The larger the bonemarrow registry and the quicker a donor can be identified, the better thechances for a successful transplant. Next-generation sequencing (NGS)
protocols are essential in routine HLA typing due to their accuracy, cost,
and throughput.1, 2 To maximize the throughput of manual NGS library
preparation, the 96-channel Gilson PLATEMASTER® pipetting system canbe easily integrated into existing workflows.

Time8-channel pipette
1104 sec

Long range PCR

1296 sec

PCR-Pooling

StepTime
PLATEMASTER
204 sec
515 sec

Purification
Fragmentation

184 sec

34 secPurification

METHODS

DNA-End-Repair

184 sec

34 secAdapter Ligation

In this experiment, the accuracy, reproducibility, and total pipetting timerequired to complete an established HLA NGS workflow was compared
between the Gilson PLATEMASTER and an 8-channel pipette.

Purification
Index-PCR

184 sec

34 sec

IPCR-Pooling

ACCURACY AND REPRODUCIBILITY

Quantification

184 sec

34 secLibrary-Pool

B

Accuracy
8-channel pipette10

measured volume [µl]86420C

25target volume [µl]10Reproducibility
8-channel pipette

total pipetting time

14 min, 15 sec

192

pipetting steps

168642025target volume [µl]10

Figure 2NGS workflow scheme (192 samples, 6 loci). Required
time for pipetting steps (bold) was compared using an
8-channel pipette (left) or the PLATEMASTER® (right).
Using PLATEMASTER® reduced the number of pipetting
steps by a factor of 12, and decreased time spent
pipetting by 73%, when compared to an 8-channelpipette.

Reproducibility
PLATEMASTER

Pipetting Time800

target vol.
mean

mean
1.5
1.0
0.5
0.0

52 min, 16 sec

10D
measured vol. [µl]measured vol. [µl]target vol.

Accuracy
PLATEMASTER

1 2 3 4 5 6 7 8 9 10 11 12column1.51.00.50.0

8-channel pipette
PLATEMASTER

600time (sec)measured volume [µl]AMiSeq

1 2 3 4 5 6 7 8 9 10 11 12column

40020001

number of plates

6

Figure 1Pipetting accuracy was measured gravimetrically at three separate
volumes in triplicate with the 8-channel pipette (A) and PLATEMASTER®
(B). Pipetting reproducibility was tested by measuring a transferred
volume of 2 µL across 12 columns of a 96-well plate.

Figure 3
The time spent preparing one and six 96-well plates
using an 8-channel pipette and PLATEMASTER®.

CONCLUSIONREFERENCESIn NGS library preparation workflows, when compared
to an 8-channel pipette, PLATEMASTER:• Maintains a high degree of accuracy
and reproducibility• Saves a significant amount of time
• Reduces tedious pipetting steps bya factor of 12, minimizing any
possibility of cross-contamination
PLATEMASTER provides
the highest throughput possible,
without the use of potentiallyexpensive robotic devices

PSTR1-01 | © 12/2017 Gilson International BV Germany. All rights reserved. All trademarks are the property of Gilson Inc and its subsidaries.

Lan, J. H. and Zhang Q.. Clinical application of next-generation sequencing inhistocompatibility and transplantation. Curr Opin Organ Transplant, 20, 461 – 467 (2015).
Chapman, M., Warren, E. H. III, Wu, C. J., Application of Next Generation Sequencing to
Blood and Marrow Transplantation. Biol Blood Marrow Transplant, 18, S151 – S160 (2012).

For further information
please contact your local
Gilson office.
www.gilson.com/ContactUs