SpeedCycler

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SpeedCycler
rapid-PCR Peltier-Thermal-Cycler
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(Available) Rapid-PCR Thermocyclers
DNA amplification by PCR can be performed
rapidly. Rapid temperature cycling for PCR is
defined as completing 30 cycles of amplification
in lt 30min Witwer et al., 1994
gt 8 C/sec 20 sec/cycle lt30min/PCR-Exp.

• Advantages
• -High speed
• -Fast temperature changes - high
• accuracy
• Disadvantages of hot air Rapid cyclers
• cappilary sample tubes
• sample preparation takes a lot of time
• unsuitable for automation

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Standard and Rapid-PCR in comparison

Standard-PCR
Rapid-PCR
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Thermalcycler products
Standard Cycler - Mostly based on Peltier
technology
Rapid Cycler
- Not using standard format
capillary-hot-air Cycler (Idahotech Roche) -
Idahotech 48 capillaries - Roche (Idahotech
license) cycler 32 capillaries
Personal Thermocycler - (16 - 48-sample
block) Classic Cycler - (96 - 1534 sample
block)
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The SpeedCycler - Rapid PCR in a Microtiter
plate

PCR Speed is always a combination between Cycler
and used Consumable
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SAC Technology (Self Adapting Container)
Small volume head space
Close up .....
Sample Volume (max. 20µl)
Low profile sample block
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SAC Technology (Self Adapting Container)
Before thermal cycling
During thermal cycling

• During thermal cycling the pressure in the head
  space rises
• The elastic walls of the ultrathin wells deform
  during process
• Container fits exactly to geometry of Sample
  Block
• Therefore Effectiveness of energy transfer higher
  than 90

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SpeedCycler in Facts

• The new standard in Quality and Speed
• Ultra fast, up to 10times faster
• Heating-Rate 10C/s and Cooling-Rate 6C/s
• Less then 13min for 30 Cycles
• Ultra thin MTP with SAC-Technology
• Efficiency of energy transfer better then 90
• 36 and 96 Well MTP available
• Clearly Higher Quality of PCR Products
• Less Primer-Mismatching
• SPS Sample Protection System
• Easy Handling
• Low noise
• Stand Alone or PC controlled
• FDA conform

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Why are other Thermocyclers so slow?
Thermal cyclers generally use microcentrifuge
tubes or modified microtiter plates. These
plastic containers insulate the sample causing
slow temperature transitions. The temperature of
a 100 µl sample within a microcentrifuge tube
lags 20-40 s. behind the block temperature. This,
combined with sluggish temperature ramp rates of
heating blocks, explains why they are so slow.
There is another major factor - temperature hold
times. A common protocol for temperature cycling
is 1 min at 94C, 2 min at 55C and 3 min at
72C. Even if instantaneous temperature
transitions were possible, one cycle would take 6
minutes. The actual enzymatic extension occurs in
about 10 seconds for a 500 bp fragment. This
leaves 97 of the total time wasted, not even
counting ramp time. What are these machines
doing? They are waiting for steady-state
conditions to bring all the samples to roughly
the same temperature. Unfortunately, because of
the geometry of the heating elements and metal
blocks, even the steady-state condition may not
produce uniform temperatures across all samples.
At best, these machines suffer from temperature
imbalance during the long wait for equilibrium
conditions. At worst, the imbalance is
permanent. While other manufacturers have
partially addressed these issues, the RapidCycler
is the only thermal cycler fast enough to
approach the kinetic limits of amplification
reactions.
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Summary
Rapid and efficient amplification of different
DNA-templates in the range of 100 bp up to 24
kb Enormous time savings by using SpeedCycler in
amplification of 100-500 bp, of 4-5 kb and 24 kb
PCR time savings gt 40 in competition to other
cyclers Clearly higher specificity of PCR-results
by using SpeedCycler caused by high accuracy of
annealing
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Rapid heat block Thermalcycler
International ApplicationInternal Appl. No
2605-PCTPCT No PCT/EP00/03224PCT Filed
05.04.2000Priority Date 08.04.1999Publicat
ion Date 19.10.2000PCT Pub. No. WO
00/61797State of application national /
regional phase
US-PatentInternal Appl. No 2605-US Appl.
No. 09/719,125Filed 07.12.2000 Priority
Date 08.04.1999 Publication Date
29.04.2003 Publication No. 6,556,940 B1
State of application Certificate
European ApplicationInternal Appl. No
2605-EP Appl. No. 99 952 199.2 Filed
05.04.2000 Priority Date 08.04.1999
Publication Date 19.10.2000 Publication No.
1 090 141 A1 State of application
Examination
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Ultrathin-walled multiwell plate for heat block
thermocycling
International ApplicationInternal Appl. No
2604-PCTPCT No PCT/EP99/08178PCT Filed
28.10.1999Priority Date 29.10.1998Publicat
ion Date 11.05.2000PCT Pub. No. WO
00/25920State of application national /
regional phase
US-ApplicationInternal Appl. No 2604-US
Appl. No. 09/830,511Filed 26.04.2001
Priority Date 29.10.1998 State of
application Examination
European ApplicationInternal Appl. No
2604-EP Appl. No. 99 952 630 Filed
28.10.1999 Priority Date 29.10.1998
Publication Date 19.09.2001 Publication
No. 1 133 359 A1State of application
Examiniation finished grant of patent (
Regel 51 (4) AO for EPÜ)