| Run |
Date and Time
|
Objective |
Attendance |
| 1 |
8 AM Sat, 9/1, 07 |
- (RBS + cinR) + (RBS + HSL) = cinR + HSL
- RBS + cinR ver. 2
- plate culture for (cinR + HSL) and (RBS + CinR) ver.2
|
茹, 翔 |
| 2 |
8 AM Sun, 9/2, 07 |
|
雄, 威, 茹, 駿 |
| 3 |
8 AM Mon, 9/3, 07 |
- plasmid extraction for (cinR + HSL) and (RBS + CinR) ver.2
- pOmpC + (RBS + INS_A)
- (RBS + cinR) + (RBS + HSL) + D-term
|
宥, 君, 威 |
| 4 |
8 AM Tue, 9/4, 07 |
- plate culture for ...
- Measure the concentration of all excised parts
- (cinR + HSL)
- (RBS + CinR) ver.2
- BsaBI digestion of cinR
|
宥, 君, 威 |
| 5 |
8 AM Wed, 9/5, 07 |
- (RBS + INS_A), PCR #1
- (RBS + INS_B), PCR #1
- pOmpC + (RBS + INS_A), not check yet
- pOmpC + (RBS + INS_B), not check yet
|
威, 茹 |
| 6 |
8 AM Thr, 9/6, 07 |
|
威 |
| 7 |
8 AM Fri, 9/7, 07 |
|
儒, 威, 茹 |
| 8 |
8 AM Sat, 9/8, 07 |
- (RBS+INS_A), PCR #2
- (RBS+INS_B), PCR #2
- GEL check (5 tubes for each gene)
- INS_A has two tubes A1 (0.08g), A3 (0.15g)
- INS_B has five tubes B1 (0.12g), B2 (0.12g), B3 (0.16g), B4 (0.11g), B5 (0.17g)
- ligation for INS_A and pOmpC
- plate culture for (pOmpC+INS_A and pOmpC+INS_B), transformation failed
|
倫, 茹 |
| 9 |
8 AM Sun, 9/9, 07 |
- plate culture failed (possible due to absent of vector, checked by text paper record)
- re-ligate INS_A insert into vector with pOmpC
- re-check the concentration of insert INS_A by [http://rsb.info.nih.gov/ij/ ImageJ]
- plate culture again for pOmpC+INS_A
|
倫, 翔 |
| 10 |
4 PM Mon, 9/10, 07 |
- plate culture is still failed
- re-build pOmpC + INS_A in two different insert concentration
- 2uL insert + 1uL vector
- 3uL insert + 1uL vector
|
倫, 翔, 銘 |
| 11 |
6 PM Thu, 9/11, 07 |
- re-measure the concentration of insert by biophotometer
- dsDNA use A260 OD to estimate, 1 A260 OD = 50 mg/mL = 0.05 ug/uL
- OD ranged from 0.1 - 2.0 is located in linear region
- OD close to zero is not reliable
- test for D7 and D8 (9/8 PCR #2) in box #1
|
銘 |
| 12 |
6 PM Wed, 9/12, 07 |
- check (9/5 PCR) by GEL separation
- concentration of INS_A is still too low
- RE-examine the condition of PCR in different annealing temperature
- PCR INS_A in 49.8, 52.3, 55.5, 58.7 and 60.9 degree
|
銘 |
| 13 |
6 PM Thur, 9/13, 07 |
- check (9/12 PCR) by GEL separation
- band seems O.K.
- best annealing temperature is around 55.5
- biophotometer concentration test
- 9/5 and 9/12 PCR seem O.K.
- 9/8 PCR seem problemistic due to A260/A280 more than 10
|
銘 |
| 14 |
6 PM Sun, 9/16, 07 |
- digest CinR+HSL (ES) and D-term (EX)and GEL separation check
- tube #1 and #2 of CinR+HSL plasmid extraction is not correct after GEL check
- first digestion of D-term (EcoRI) is not very sucessful
- thus pett re-digest D-term with double enzyme amount
|
宥, 君, 翔 |
| PCR |
Thur, 9/27, 07 |
- PCR of TAT_INS_A, TAT_INS_B, OmpRBS, TAT_IDE
|
威 |
