If you are checking wheather it exists then change to this code.
When youu use greater than then it means even if the record are more than one the result with still be the same
If ds.Tables("Job_card_opening").Rows.Count > 0 Then
ins = "insert into Job_card_opening (UserName,Designation,Barcode_number,IN_Time,Transaction_Completed) values ('" & TextBox1.Text & "','" & TextBox2.Text & "','" & TextBox3.Text & "','" & Now & "','0')"
Please comment your code for ease understanding