gDNA

The cDNA was synthesized according to PrimeScript RT reagent Kit with gDNA Eraser instruction, which produced by Primers Design TaKaRa Bioengineering (DaLia n) Co., Ltd and stored at -20AdegC before using.

coli was performed with Thermo ScientificA(r) GeneJET Genomic DNA (gDNA) Purification Kit# K0721 by using manufacturer's protocol.

Trizol reagent, primescript RT reagent kit with gDNA eraser and SYBR premix Ex Taq kit were purchased from Takara (Dalian, China).

Genomic deoxyribonucleic acid (gDNA) was extracted from peripheral blood leukocytes in whole blood samples using QIAamp DNA Mini Kit (QIAGEN, Hilden, Germany) following the manufacturer's instructions.

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The testing process used during this verification consisted of 2 steps: manual specimen preparation to obtain genomic DNA (gDNA) from formalin-fixed, paraffin-embedded (FFPE) tissue or plasma specimens using the cobas DNA Sample Preparation Kit (Roche Diagnostics) or cobas cfDNA Sample Preparation Kit (Roche Diagnostics), followed by PCR amplification and detection of targeted KRAS mutations using the KRAS Mutation Test v2 kit (Roche Diagnostics; Figure).

Samples were stored at 4 [degrees]C until genomic DNA (gDNA) extraction.

Approximately 3ml of blood was collected in a Vacutainer Potassium-salt EDTA tube based on the protocol from the ReliaPrep[TM] Blood gDNA Miniprep System (Promega, USA, Cat# A5081).

In brief, after centrifugation, the cell suspension was filtrated from the gDNA eliminator column, then transferred and attached to the RNeasy spin column and washed with the solutions as instructed.

The gDNA (genomic DNA) was extracted from Jatropha cinerea leaves that were collected during the seed collection and stored at -80[degrees]C prior to use.

The cDNAs (10 [micro]l) were synthesized from 2 [micro]g RNA of each sample by utilizing reverse transcription kit (PrimeScript™ RT reagent Kit with gDNA Eraser RR047Q, Takara).

Genomic DNA (gDNA) was isolated from peripheral blood samples of patients according to QIAamp Blood kit protocol (Qiagen, Hilden, Germany).