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Grasses are the most commonly encountered botanical evidence in outdoors crime scene. Being ubiquitous in nature the grasses tend to serve as the best contact and trace evidence during a forensic investigation. Historically, these samples were identified using morphological techniques, which work well if the evidence is intact. Grasses/botanical samples are generally found in degraded, dried and contaminated forms. These types of samples lead to scarcity of vital morphological characteristics that are necessary for its correct identification. Therefore, molecular identification plays an important role in the accurate identification of the degraded and dried botanical evidence. The present study was designed as a preliminary effort in the amplification of the ITS 2 marker from the DNA extracted from chemically treated grass stains. 10 grass species were taken for the present study from six subfamilies. The stains were produced on the cotton cloth piece and were further treated with the cleaning agents (Bleach, NaOH, Soap and Gasoline). Seven out of 10 species were successfully amplified with 330-370 bp amplicons leading to 70% PCR efficiency rate. In the current study, the effect of chemicals on grass stained cotton cloth piece was studied and its impact on the recovery and amplification of ITS 2 marker.