Red chili (Capsicum annum L.) is a horticultural commodity that is widely consumed by Indonesian people. The need for red chili is increasing along with population growth. One of the efforts to increase red chili production is fertilization. This study aimed  to determine the effect of the dose of manure and the frequency of application of biological fertilizers on the growth and yield of red chili. This research was conducted in May 2021-September 2021, at the Screen House 3rd floor of Building C, Faculty of Agriculture, Universitas Sebelas Maret, Surakarta. The method used was a Randomized Completely Block Design (RCBD) which was arranged in a factorial manner consisting of two treatment factors and three replications. The first factor was the ratio of soil and manure (without manure, 3:1, 3:2, and 3:3). The second factor was the frequency of application of biofertilizer (without biological, once in 14 and once in 28 days). Data analysis used variance at a level of 5% and continued with Duncan’s Multiple Range Test at a level of 5% if there was a significantly different effect. The results showed that the ratio of soil and manure (3:2) had an effect on increasing the number of leaves. The frequency of giving biofertilizer treatment once every 14 days had an effect on increasing the number of leaves and stem diameter. There was no interaction between combination the dose of manure and the frequency of biofertilizer application on the growth and yield of red chili.