A reproducible regeneration protocol for ICGV 12991, CG 7 and Red Valencia groundnut genotypes using Cotyledonary Node explants has been optimized. The effect of different BAP concentrations combined with either 2,4-D or TDZ was tested to determine optimum conditions for high shoot induction. Different BAP concentrations were tested to determine an optimum concentration for shoot elongation. Different NAA concentrations were similarly evaluated to determine the best concentration for rooting. Media containing combination of 5 mg/L BAP and 1 mg/L TDZ was the best concentration for shoot induction while media containing BAP at 5 mg/L was the best for elongation of shoots. NAA concentration of 1 mg/L gave the highest number of plants with roots. This works provides a very good protocol which will be beneficial during groundnut tissue culture as well as genetic transformation of groundnuts.