<h3>ABSTRACT</h3> <h3>Background</h3> One major challenge for detecting the virus that causes COVID-19 is commercial SARS-CoV-2 testing kit or reagent availability. To allow every laboratory or hospital access to an in-house assay, we developed two low-cost SARS-CoV-2 detection assay protocols using in-house primers and reagents/equipment on hand in most biology or diagnostic laboratories: a SYBR Green–based RT-PCR and PCR assays. RNA extraction has also become a major bottleneck due to limited supplies and the required labor. Thus, we validated alternative RNA extraction protocols. <h3>Methods</h3> SARS-CoV-2 genome sequences deposited into the GISAID database were retrieved to design and synthesize in-house primers. Forty patient samples were collected by nasopharyngeal swab, coded, and used to develop and validate the assay protocols. Both assays used TRIzol and heat-processing techniques to extract RNA from patient samples and to inactivate the virus; thus, testing was conducted in a conventional biosafety level 2 laboratory. <h3>Results</h3> The sensitivity and specificity of the primers were evaluated using samples previously confirmed positive for SARS-CoV-2. The positive amplicons were sequenced to confirm the results. The assay protocols were developed, and the specificity of each PCR product was confirmed using melting curve analyses. The most accurate heat-processing technique for primers with short amplicon lengths was 95 °C for 15 mins. Of 40 samples, both the SYBR Green–based quantitative RT-PCR assay and the PCR assay detected SARS-CoV-2 target genes in 28 samples, with no false-positive or false-negative results. These findings were 100% concordant with those of the diagnostic laboratory that tested the same samples using a Rotor-Gene PCR cycler with an Altona Diagnostics SARS-CoV-2 kit (R2=0.889). <h3>Conclusions</h3> These approaches are reliable, repeatable, specific, sensitive, simple, and low-cost tools for the detection of SARS-CoV-2 in a conventional biosafety level 2 laboratory, offering alternative approaches when commercial kits are unavailable or cost ineffective.