K-RAS 1.4

LCD-Array Kit

  • Fast
    120 min PCR / 45 min array protocol / 5 min analysis Fast First results after ~3 hours  (excl. sample preparation)

 

  • Sensitive
    Detects less than 0.5 % mutations in wild type background (clamping PCR)
    170 bp amplicon suited for FFPE material

 

  • Specific
    6 point mutations in codon 12
    3 point mutations in codon 13

 

  • Robust
    Non-fluorescent detection for easy read-out and long term storage
    Fully automated read-out by SlideReader software, minimal instrumentation

 

 

Product Information

Specificity

Fast and reliable detection of nine point mutations within codon 12 & 13 of the k-ras oncogene. All nine mutations lead to amino acid changes and have been reported to hamper antibody based therapies.

Assay Principle

Using extracted human DNA from micro dissections as template, biotinylated PCR amplicons (170 bp) are generated in a single reaction. To increase the sensitivity, reactions can be performed in the presence of a ‘Wild-type’ Suppressor Compound (WSC), which selectively suppresses the amplification of wild type sequences. The labeled amplicons are hybridized to sequence specific capture probes immobilized on the LCD-Chip surface. Following a short wash routine, comprising high stringency, visualization of bound amplicons is mediated by an enzyme-substrate cascade. The LCDChip contains eight identical micro arrays separated in small reaction chambers. Each array can be addressed individually, allowing the parallel analysis of eight samples on one chip. Following PCR amplification the complete procedure takes 45 minutes. Data analysis can be achieved using the SlideReader software.

 

Array Pattern

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Fig.1:

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Parallel analysis of eight samples on one LCD-Chip. The
array field spacing is compatible with standard multi pipettes.

Fig.2 :
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Hybridizations of three different samples to K-RAS 1.4 arrays
A) Wild type sample, no mutations detectable
B) Codon 13 Gly>Cys mutation within a wild type background
C) Codon 12 Gly>Ser mutation within a wild type background

 

* The Polymerase Chain Reaction (PCR) process is covered by U.S. patents owned by Hoffmann-La Roche, Inc. and patents owned by F. Hoffmann-La Roche Ltd. No license under these patents to use the PCR process is conveyed expressly or by implication to the purchaser by the purchase of this product.

 

Software

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SlideReader Vers. 8.01

• Fully automated image analysis
• comprehensive data reports in HTML format
• fast & intuitive navigation
• multiple user accounts

 

Instrumentation

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CHIP-Scanner PF2700

• Transmission light scanner for LCD-Arrays,
• 10 µm resolution
• 16 bit grey scale TIFF images

CHIP Spin FVL2400

• Bench Top mini centrifuge
• 2400 rpm, constant
• Adaptor for LCD-Arrays
• incl. 3 rotors

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