HTS Just Got Better

Assay for Endogenous Gene Expression

LightCycler photo
As compared to Reporter Gene Assays, qPCR enables you to:
  • Obtain more biologically relevant data
  • Increase data sensitivity
  • Decrease number of false hits
LightCycler logo

Replace Reporter Gene Assays

Watch to Learn More

SLAS 2015: In the video above, Dr. Rupal Cutting explains how Roche’s new high-throughput, completely automatable qPCR workflow easily surpasses reporter gene assays.

Known Issues

Reporter Gene Assays

Unlike reporter gene assays, RT-qPCR can be used on native, non-transformed cell types, and RT-qPCR measures mRNA transcripts of the actual gene as it is expressed from within its natural environment, rather than transcripts from an artificial, plasmid-based construct.*

Taken from LabCyte White Paper, “High Throughput RT-qPCR for Gene Expression Analysis.”

Cited Papers Referencing Known Issues:

  1. Mechanism of PTC124 activity in cell-based luciferase assays of nonsense codon suppression.

    Douglas S. Auld, Natasha Thome, William F. Maguire, and James Inglese. NIH Chemical Genomics Center, National Institutes of Health, Bethesda, MA 20892-3370

  2. False Positives in a Reporter Gene Assay: Identification and Synthesis of Substitutes N-Pyridin-2-yl-benzamides as Competitive Inhibitors of Firefly Luciferase.

    J Med Chem, 2008, 51, 4727-4729 Heitman, L.H.; Veldhoven, J.P.D.; Zweemer, A.M.; Ye, K.; Brusse, J.; Ijzeman, A.P.

Time for a Change?

Disadvantages of Reporter-Based Screening

Lengthy Time-to-Completion

Elevated False Positives & Negatives

Less Biologically Relevant

Shorten Your Path

Go Directly from your GOI into qPCR Screen

Reporter gene assays

"Identify GOI → Clone → Sequence → Transfect → Establish stable cell line/transient → transfection → Over express GOI → Record GOI levels → Validate using qPCR"

qPCR screen

From Cells to Results
with No Manual Intervention

Fully Automatable Workflow
1 Start with treated cells in a 384- or 1536- well tissue culture plate
2 Add lysis buffer, incubate at room temperature for 5 minutes
3 Transfer lysates and one-step master mix to qPCR plate
4 Seal and centrifuge
5 Amplify (RT-qPCR)

Real Versatility

Interface with a Wide Array of Liquid and Plate Handlers

Plate Handlers

  • Labcyte Access Laboratory Workstation
  • Thermo Scientific Orbitor RS
  • HighRes Biosolutions Plate Valet

Liquid Handlers

  • Labcyte Echo® 555
  • EDC Biosystems ATS-100
  • BioDot AD1500

LightCycler® 1536 Details

The LightCycler® 1536 optical unit facilitates mono- and dual-color assays using a variety of dyes, such as FAM, VIC, and Yellow 555, by supporting formats including Universal ProbeLibrary (UPL) probes, hydrolysis probes, and intercalating dyes (e.g., BrightGreen). The unique thermal block cycler design, in combination with the LightCycler® 1536 Multiwell Plate, enables very high well-to-well homogeneity and inter-assay reproducibility.

The LightCycler® 1536 Software supports the next generation of real-time PCR results (e.g., Cp's, endpoint fluorescence values), providing novel threshold functions for data refinement and powerful tools for data sorting and reviewing. Since this system is designed for automated high-throughput laboratory workflows, the instrument’s software allows easy data management for network and LIMS environments. In combination with Roche RealTime ready reagents, the system also features a new surveillance concept that checks for pipetting errors.

Instrument Specifications
(without computer)
57.4 cm x 58.8 cm x 49.7 cm
22.6 in x 23.1 in x 19.6 in
(W x D x H)
Weight 55.6 kg (121 lbs)
Computer PentiumPC with Windows XP
Sample Number 1536
Reaction Vessel LightCycler® 1536 Multiwell Plate
Reaction Volume 0.5 – 2.0 µl
Excitation Source Xenon lamp
Filters Excitation: 465 nm, 533 nm
Detection: 510 nm, 580 nm
Detection Cooled monochrome CCD camera
Temperature Range +37to +95°C
Heating Rate 4.8°C/sec
Cooling Rate 2.5°C/sec
Temperature Control Peltier-based heating/cooling
Therma-BaseTM heat spreading technology
Run Time <50 minutes
Multiwell Plate Thermaxis® technology (two-component design)
Barcode labeled for automated workflow tracking
  • User-friendly programming
  • Compact overview of basic results (e.g., CPs, slopes, endpoint fluorescence), and surveillance data
  • Novel threshold functions for data refinement
  • Powerful tools for data sorting and reviewing
  • Multiple data input/output formats
  • Simple data management for network environments
  • Result data accessible via LIMS
  • Automation integration module (AIM) for full connectivity to both remotely control the LightCycler® 1536 System and to exchange experiment data with other software
  • Software Development Kit to establish the remote application between the LightCycler® 1536 System and other instrumentation

Interested in speaking with us?

Additional Resources

Product Details

The Changing Face of HTS - White Paper

The Future of High Throughput Assays Webinar

iPS cell derived neuronal cells for drug discovery

High Throughput RT-qPCR for Gene Expression Analysis White Paper

Video: 1536 qPCR- A Robust Method for Detecting Changes in Gene Transcript
PJ Aspesi, Scientist at Novartis Institutes for BioMedical Research

Video: Thermal Shift Assays on LightCyclers
Rama Badugu, Ph.D., Application Support Consultant at Roche Diagnostics

Video: Enabling High Throughput qPCR
Lynn Rasmussen, HTS Supervisor at Southern Research Institute

Video: High Throughput RT-qPCR for Identifying Activators of Latent HIV-1 in Primary Human CD4+ T Cells
Helen Yu, PhD, Biology Department, Gilead Sciences

Video: Validation of an automation compatible high-throughput Reverse Transcription qPCR workflow
Dr. Catherine Bardelle, AstraZeneca, Alderley Park, Macclesfield, UK

Speak with a scientist

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