Rotor Gene 6000 Real-Time PCR Machine Features: • Unique centrifugal rotary format • Uniformity: ± 0.01°C • Resolution: 0.02°C (smallest programmable and reported thermal increment) • 5-Plex (Green, Yellow, Orange, Red, Crimson) fluorescence channels • High resolution melt (HRM) application • Holds 36 x0.2ml tubes, 72 x 0.1ml strip tubes or 100 Gene-Disc • Software: Build-in extensive analysis, graphing and statistical functions. Unlimited use software license for Windows XP, Pentium IV (2GHz) or higher PC. Rotor Gene 3000 Real-Time PCR Machine Features: • Unique centrifugal rotary format • Excitation Source: LED high power diodes, 470nm, 530nm, 585nm and 625nm • Detection Filters: 510nm (FAM), 555nm (JOE), 610nm (ROX) and 660nm high-pass (Cy5) • Holds 36 x0.2ml tubes or 72 x 0.1ml strip tubes • Software: Build-in extensive analysis, graphing and statistical functions. Use of the machines in the Molecular Biology Facility • The Rotor-Gene machines are in high demand therefore there is an (external website). Plan ahead, book time slots and stick to them. • Users are responsible for supplying all their own plastic ware and reagents.

The QIAsymphony® AS is ideal for laboratories performing routine PCR tests on a. The comprehensive Rotor-Gene Q software package supports all current.

0.1ml tubes and Gene-Disc 100 are produced by Corbett especially for the Rotor-Gene. • Save files in your own folder in the 'Rotor Gene Run File' folder in the local disk (C:). The computer is also linked to the network for easy transfer of your data to the server. Tips and addition information • The Rotor-Gene machines support reaction volumes from 5 µl to 100 µl (rotor and protocol dependent).

Typical reaction volume is 20µl. • The use of a normalisation dye, such as ROX reference dye, is optional on the Rotor-Gene. • Many companies supply master mixes for real-time PCR which can decrease the chance of pipetting errors in this highly sensitive method. • If you would like to learn more about the real-time PCR experimental design and data analysis, please contact Dr. Donna Lai, the Molecular Biology Officer. Bioanalyzer, Real-time RT-PCR workshops are conducted occasionally during the year.

Please contact Dr. Donna Lai for details.

Unique rotary design for outstanding performance The unique centrifugal rotary design of the Rotor-Gene Q makes it the most precise and versatile real-time PCR cycler currently available (see figure 'Cross-section of the Rotor-Gene Q'). Each tube spins in a chamber of moving air, keeping all samples at precisely the same temperature during rapid thermal cycling. Detection is similarly uniform. When each tube aligns with the detection optics, the sample is illuminated and the fluorescent signal is rapidly collected from a single, short optical pathway.

This thermal and optical uniformity results in sensitive, precise, and fast real-time PCR analysis (see figure 'Precise real-time PCR analysis'). It also eliminates sample-to-sample variations and edge effects. These are unavoidable in traditional block-based instruments due to temperature gradients across the block and multiple, complex optical pathways.

The rotary design delivers: • Well-to-well variation ±0.02°C • Uniform detection eliminating the need for ROX reference dye • Fast ramping and negligible equilibration times for short run times • Complete confidence in your results! Unrivaled optical range enables multiple applications Whether your assay is based on intercalating dyes such as SYBR Green, probes such as hydrolysis (TaqMan), hybridization (FRET), Scorpion probes, or other multiplex chemistries, the Rotor-Gene Q meets your requirements. With up to 6 channels spanning UV to infrared wavelengths, the cycler delivers the widest optical range currently available (see Table 'Channels for optical detection'). In addition, the software allows you to create new excitation/detection wavelength combinations, which means that the Rotor-Gene Q is compatible with dyes you may use in the future. Expand your research with HRM High-resolution melting analysis (HRM) is a closed-tube, post-PCR analysis that has raised enormous scientific interest.

HRM characterizes double-stranded PCR products based on their dissociation (melting) behavior. It is similar to classical melting curve analysis, but provides far more information for a wider range of applications.

PCR products can be discriminated according to sequence, length, GC content, or strand complementarity, down to single base-pair changes. Previously unknown and even complex sequence variations can be readily detected and characterized in a robust and straightforward way. The rotary design of the Rotor-Gene Q and its outstanding thermal and optical performance are highly suited to HRM. The HRM option for the Rotor-Gene Q includes: • A specially tuned high-intensity optical HRM channel • Thermal resolution down to 0.02°C • High data acquisition rates • Comprehensive HRM software The Rotor-Gene Q is the only real-time cycler currently capable of deciphering the most difficult class IV SNPs by HRM. Harness the power of HRM using dedicated QIAGEN HRM Kits for applications such as genotyping (see figure 'HRM analysis of a class IV SNP with less than 0.1˚C difference between homozygote alleles' for data from the Type-it HRM PCR Kit), quantitative methylation analysis (see figure 'Highly sensitive results with detection of even low percentages of methylated DNA' for data from the EpiTect HRM PCR Kit), gene scanning, and sequence matching. The Type-it HRM PCR Kit reliably and accurately detects gene mutations and SNPs.