Use the query :
Select * from Table_name WHERE Column_Name = Your_Value
Lets say your result datatable has 5 rows and you want to bind Column1 to your dropdown.
All you need to do is just bind your values from datatable to dropdownlist
Here Iam changing your code
DataTable dt1 = new DataTable();
SqlConnection con1 = new SqlConnection(str);
con1.Open();
string query1 = "select Lot_No from tblStockDetail where ItemID = '" + txtItemID.Text + "'";
SqlDataAdapter ad1 = new SqlDataAdapter(query1, con1);
ad1.Fill(dt1);
cboLotNo.DataSource = dt1;
cboLotNo.DataTextField = "Column_Name";
cboLotNo.DataValueField = "Column_Name";
cboLotNo.DataBind();
txtQty.Focus();