NYMU Taipei/Lab Notes/2007 9 11
From 2007.igem.org
(Difference between revisions)
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** two different PCR (INS_A and INS_B) have been done by 海威 (9/5, 2007) and 惇茹 (9/8, 2007) | ** 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 | ** we had failed three times for construction of pOmpC + INS_A using 惇茹's INS_A and following three different ligation protocol | ||
| - | *** insert (1 uL) vs. vector (1 uL) at 09/09, 2007 | + | *** insert is INS_A and vector is pOmpC |
| - | *** insert (2 uL) vs. vector (1 uL) at 09/10, 2007 | + | *** 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 (3 uL) vs. vector (1 uL) at 09/10, 2007 | + | *** 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 | *** a trivial failure (absent of vector part) at 09/08, 2007 | ||
** According to YEA ligation protocol, it requires | ** According to YEA ligation protocol, it requires | ||
Revision as of 15:06, 11 September 2007
- 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
