From 2007.igem.org

• Fresh back
  • two different PCR (INS_A and INS_B) have been done by 海威 (9/5, 2007) and 惇茹 (9/8, 2007)
  • we had failed three times for construction of pOmpC + INS_A using 惇茹's INS_A and following three different ligation protocol
    • insert is INS_A and vector is pOmpC
    • insert (1 uL) vs. vector (1 uL) at 09/09, 2007: the quantity are 6ug = 6,000 ng vs. 13ug = 13,000 ng (original measure)
    • insert (2 uL) vs. vector (1 uL) at 09/10, 2007: the quantity are 12ug = 12,000 ng vs. 13ug = 13,000 ng (original measure)
    • insert (3 uL) vs. vector (1 uL) at 09/10, 2007: the quantity are 18ug = 18,000 ng vs. 13ug = 13,000 ng (original measure)
    • a trivial failure (absent of vector part) at 09/08, 2007
  • According to YEA ligation protocol, it requires
    • ligation buffer A 1uL
    • ligation buffer B 1uL
    • vector 50 ng or 100 ng (proposed by pett)
    • insert 300 ng
    • ligase 1uL
    • add water to make total volume as 10 uL