877.777.1874 • www.asuragen.com

*�Research�Use�Only.�Not�for�use�in�diagnostic�procedures.

Multiplex assay for 7 distinct mutations in KRAS codon 12 and 13

Co-detection of a KRAS endogenous control sequence in the same reaction

Positive and Negative control samples provided independently

No cross reactivity between mutations and automatic detection on the Luminex® 100 ISTM or 200TM systems

Simultaneous detection and identification in a single reaction well of a 96-well plate

Assess DNA quality and integrity in each DNA specimen

Assess validity of the PCR, hybridization and detection steps in each run

Easy to interpret qualitative results

KRAS�is�an�oncogene�involved�in�the�epidermal�growth�factor�receptor�(EGFR)�signaling�pathway�that�controls�cancer�cell�proliferation�and�apoptosis.�Recently,�there�has�been�increased�interest�in�the�use�of�specific�biomarkers�in�colorectal�cancer�(CRC)�and�other�cancers�treated�with�EGFR�targeted�therapies.�The�Signature®�KRAS�Mutations�7�(RUO)*�assay�is�based�on�the�Signature®�Technology�Platform�for�the�rapid�multiplex�analysis�of�nucleic�acid�sequences,�allowing�detection�of�up�to�100�DNA�or�RNA�targets�in�a�single�reaction�well.�The�Signature®�KRAS�Mutations�(RUO)*�provides�comprehensive�information�on�7�distinct�mutations�within�codons�12�and�13�of�the�KRAS�oncogene.�The�96-well�assay�format�enables�characterization�of�KRAS�mutation�status�in�approximately�4�hours�using�a�broad�input�range�of�genomic�DNA�isolated�from�cultured�cells�or�fresh,�frozen�or�formalin-fixed�paraffin-embedded�(FFPE)�specimens.�The�streamlined�workflow�and�multiplex�assay�format�increase�specimen�throughput,�decrease�reagents/supplies�use�and�reaction�set�up�time,�and�overall,�reduce�the�cost�per�individual�mutation�tested.

The�same�technology�is�also�available�for�detection�of�12�KRAS�mutations�and�BRAF�V600E�as�part�of�the�Signature®�KRAS/BRAF�Mutations�(RUO)*�Assay.

Signature® KRAS Mutations 7 (RUO)*

Figure 1:�Seven�distinct�KRAS�mutations�can�be�detected�by�the�assay�in�a�single�reaction�(right).�The�streamlined�assay�workflow�is�optimized�for�rapid�time�to�results�with�minimum�hands-on�time�(above).

G12S� (GGT>AGT)G12R� (GGT>CGT)G12C� (GGT>TGT)G12D� (GGT>GAT)G12A� (GGT>GCTG12V� (GGT>GTT)G13D� (GGC>GAC)

Key Attributes of Signature® KRAS Mutations 7 (RUO)*:

877.777.1874 • www.asuragen.com

2500-0220REV2

Excellent analytical specificity across all 7 mutations using the Signature® KRAS Mutations 7 Assay*

At least 1% analytical sensitivity using the Signature® KRAS Mutations 7 Assay*

Kit Negative control

Kit Positive control

No DNA Control

Cell line WT (HT-29)

Cell line G12V (HOM)

Cell line G12A (HET)

Cell line G12D (HET)

Cell line G12C (HOM)

Cell line G12S (HET)

Cell line G13D (HET)

Plasmid G12R in HT-29

G12V, GTT

117

3771

142

153

5202

135

169

186

167

240

197

G12A, GCT

106

206

258

171

315

6732

173

186

164

147

245

G12D, GAT

96

105

127

133

106

367

5317

84

125

158

184

G12C, TGT

105

15

108

126

114

128

87

5180

141

93

221

G12S, AGT

169

169

165

213

211

135

94

498

3845

189

181

G13D, GAC

162

172

139

81

83

125

135

53

112

4245

411

G12R, CGT

176

207

120

218

222

102

191

214

87

112

3537

KEC

4912

5292

118

8608

8994

8803

8656

8483

9125

8497

7986

Sample

Signature® KRAS Mutations 7 (RUO)*

Figure 2:�Representative�assay�MFI�output�data�with�10�ng�of�genomic�DNA�purified�from�7�cultured�cell�lines�either�wild�type�(WT)�for�KRAS�or�carrying�specific�homozygous�(HOM)�or�heterozygous�(HET)�KRAS�mutations.�Detection�of�G12R�mutation�target�was�tested�using�a�plasmid�DNA�carrying�the�GGT>CGT�mutation�(5,000�copies�spiked�into�10�ng�of�WT-HT-29�cell�line�DNA).�KEC�=�KRAS�Endogenous�Control.�

Figure 3:�Representative�examples�of�analytical�sensitivity�with�genomic�DNA�purified�from�a�homozygous�G12V�positive�cell�line�or�a�heterozygous�G13D�positive�cell�line.�The�DNA�samples�were�either�undiluted�(100%)�or�diluted�in�a�background�of�wild�type�genomic�DNA�(HT-29)�keeping�the�total�input�constant�at�20�ng.�A�sensitivity�equivalent�to�at�least�1%�(1�ng�of�positive�DNA�in�19�ng�of�WT�DNA)�was�achieved.

References:1�Walther�A,�Johnstone�E,�Swanton�C,�Midgley�R,�Tomlinson�I,�and�Kerr�D.�Genetic�prognostic�and�predictive�markers�in�colorectal�cancer.�Nat�Rev�Cancer.�2009�Jul;9(7):489-99.2�Linardou�H,�Dahabreh�IJ,�Bafaloukos�D,�Kosmidis�P,�and�Murray�S.�Somatic�EGFR�mutations�and�efficacy�of�tyrosine�kinase�inhibitors�in�NSCLC.�Nat�Rev�Clin�Oncol.�2009�Jun;6(6):352-66.

[P/N 49418] 48 Reactions[P/N 49419] 48 Reactions

Ordering Information Signature® KRAS Mutations 7 (RUO)*

*�Preliminary�research�data,�the�performance�characteristics�of�this�assay�are�not�yet�established.

Asuragen,�Inc.�•�2150�Woodward�St.,�Ste.�100�•�Austin,�TX�USA�78744P:�512.681.5200�•�F:�512.681.5201