Genomics Application Notes

Table 1. Processing Time

MAJOR PROCESS DESCRIPTION

AUTOMATED/ HANDS ON TIME** 24 SAMPLES

48 SAMPLES

96 SAMPLES

cDNA Synthesis: Day 1 Prepare Reagents/Set up Inst

25 min

27 min

30 min

Method Run

4 hrs

4 hrs 20 min 4 hrs 47 min

4 hrs 50 min 5 hrs 20 min

Total

4 hrs 25 min

Library Construction: Day 2 Prepare Reagents/Set up Inst

25 min

27 min

30 min

Method Run

2 hrs 30 min 2 hrs 55 min 7 hrs 20 min

2 hrs 40 min 3 hrs 7 min 7 hrs 54 min

3 hrs 5 min

Total Total

3 hrs 35 min 8 hrs 55 min

**Timing does not include thawing of reagents and PCR thermocycling

Materials REAGENTS

SOURCE

TruSeq Stranded mRNA Sample Preparation Kit

Illumina, San Diego, CA Beckman Coulter, Brea, CA Life Technologies, Carlsbad, CA

AMPure XP RiboGreen PicoGreen

Life Technologies

R6K ScreenTape

Agilent Technologies, Santa Clara, CA

MiSeq Reagent Kit v2

Illumina Illumina

TruSeq Rapid SBS 200 Cycle Kit

Tomato RNA Drosophila RNA Daphnia pulex RNA Tribolium RNA Enchenopa binotata and Umbonia crassicornis RNA

RNA samples provided by various researchers in the Department of Biology at Indiana University

Universal Human Reference RNA

Agilent Technologies Agilent Technologies

TapeStation 2200 D1K High Sensitivity Screentape with the associated reagents and consumables

LABWARE

SOURCE

Biomek Span-8 P1000 Tips, Pre-sterile with Barrier Biomek Span-8 P250 Tips, Pre-sterile with Barrier Biomek P50 Tips, Pre-sterile with Barrier Biomek AP96 P20 Tips, Pre-sterile with Barrier Biomek AP96 P250 Tips, Pre-sterile with Barrier

Beckman Coulter Beckman Coulter Beckman Coulter Beckman Coulter Beckman Coulter Beckman Coulter Beckman Coulter Beckman Coulter Beckman Coulter Beckman Coulter Fisher Scientific

Quarter Reservoir

Reservoir, Half

Quarter Reservoir, Divided by Length

Frame for Reservoirs

Agencourt® SPRIPlate® 96R - Ring Super Magnet Plate Abgene 96-Well Storage Plate, Square Well, 1.2 mL 2mL SuperClear™ Screw Cap Microcentrifuge Tubes- Conical Bottom Hard-Shell® Thin-Wall 96-Well Skirted PCR Plates

VWR

Bio-Rad Laboratories

Instruments Biomek® FX P Laboratory Automation Workstation (P/N B05450)

Illumina MiSeq

Agilent 2200 TapeStation Instrument

Illumina HiSeq 2500

Eppendorf MasterCycler ep or equivalent thermocycler with heated lid

B2013-14332 IB-18319A