Protocol detail

GAS5 SNPs Analysis for Cancer Evaluation

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Two SNPs of GAS5: rs145204276 (Ins/Del) and rs55829688 (T/C) were chosen due to their effects on lung cancer and other neoplasms [15 (link),16 (link),17 (link)]. The genotyping procedure is referred to in our previous research [18 (link)]. The genomic DNA was extracted from leukocytes in venous blood sample via QIAamp DNA kits (Qiagen, Valencia, CA, USA), and all steps with QIAamp DNA kits were in accordance with manufacture’s instruction. The reason we used leukocytes to take genomic DNA is because it is a simple way to obtain human genomic DNA. Isolated DNA was put in one refrigerator under −20 degrees Celsius. The two GAS5 SNPs were examined via the ABI StepOne Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). After that, the polymorphisms of both GAS5 SNPs were analyzed with the assistance of the TaqMan assay technique as well as the SDS version 3.0 software (Applied Biosystems) to enhance Real-Time PCR completeness.

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Hsieh M.H., Wu Y.L., Tsao T.C., Huang Y.W., Lin J.C., Lee C.Y., Hsieh M.J, & Yang S.F. (2022). Impact of LncRNA GAS5 Genetic Variants and the Epidermal Growth Factor Receptor Phenotypes on the Clinicopathological Characteristics of Lung Adenocarcinoma Patients. International Journal of Environmental Research and Public Health, 19(16), 9971.

Publication 2022

QIAamp DNA kits ABI StepOne Real-Time PCR System TaqMan assay technique SDS version 3.0 software

Assay Genomic Human genomic In blood Leukocytes Lung cancer Neoplasms Polymorphisms Real time pcr Snps Venous

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Variable analysis

independent variables

Rs145204276 (Ins/Del)
Rs55829688 (T/C)

dependent variables

Effects on lung cancer and other neoplasms

control variables

Genomic DNA extracted from leukocytes in venous blood sample
QIAamp DNA kits (Qiagen, Valencia, CA, USA) used for DNA extraction
DNA samples stored at -20 degrees Celsius
ABI StepOne Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) used for SNP genotyping
TaqMan assay technique and SDS version 3.0 software (Applied Biosystems) used for SNP analysis

controls

No positive or negative controls were explicitly mentioned in the provided information.

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