Consider a matrix M chosen uniformly at random from a class of m × n matrices of zeros and ones with prescribed row and column sums. A partially filled matrix D is a defining set for M if M is the unique member of its class that contains the entries in D. The size of a defining set is the number of filled entries. A critical set is a defining set for which the removal of any entry stops it being a defining set.For some small fixed ε > 0, we assume that n m = o(n 1+ε ), and that λ 1/2, where λ is the proportion of entries of M that equal 1. We also assume that the row sums of M do not vary by more than O(n 1/2+ε ), and that the column sums do not vary by more than O(m 1/2+ε ). Under these assumptions we show that M almost surely has no defining set of size less than λmn − O(m 7/4+ε ). It follows that M almost surely has no critical set of size more than (1 − λ)mn + O(m 7/4+ε ). Our results generalise a theorem of Cavenagh and Ramadurai, who examined the case when λ = 1/2 and n = m = 2 k for an integer k.