Difference between revisions of "Team:KAIT JAPAN/lab-note"
| Line 37: | Line 37: | ||
<dt>August 31st</dt> | <dt>August 31st</dt> | ||
<dd>PCR of ligated JAK5'+3'(extension time 3:45)</dd> | <dd>PCR of ligated JAK5'+3'(extension time 3:45)</dd> | ||
| + | <dt><font size="5" font color="#3396FF">SEPTEMBER</font></dt> | ||
<dt>September 1st</dt> | <dt>September 1st</dt> | ||
<dd>PCR ligated JAK5' and 3' (extension time 3:45) <br>diluted 100x, 1000x, 10000x restriction digests and gel purificated STAT1.<br>restriction digests and gel purification of HlyD, HlyB</dd> | <dd>PCR ligated JAK5' and 3' (extension time 3:45) <br>diluted 100x, 1000x, 10000x restriction digests and gel purificated STAT1.<br>restriction digests and gel purification of HlyD, HlyB</dd> | ||
Revision as of 07:58, 1 November 2017
LAB NOTE
- August 15th
- Presentation at Promega for sponsorship of reagents.
- August 16th
- Transformed designated parts into DH5α
BBa_I20270, R0040, J364000 ,J364001, J364002, J364003, J364004, J364005 - August 17th
- Broth culture shows no growth
- August 18th
- Transformed designated parts into DH5α
BBa_I20270, R0040, J364000, J364001, J364002, J364003, J364004, J364005
Orders from IDT
・IL-10 receptor
・STAT1
・JAK
・GFAP promotor
・HlyB
・HlyD
・IL-12 - August 19th
- Colonies were formed on all plates
But only a small number of colonies were found on the plates I20270, J364004 and J364005. - August 20th
- Open campus
- August 21st
- Transformed I20270, J364004 and J364005 into DH5α.
- August 22nd
- Precultured all 8 of the parts, then cultured them for 16 to 18 hours.
- August 23rd
- Carried out measurements for interLab study
- August 24th
- Organized the data measured
interLab study completed! - August 25th
- Received gBlocks gene fragments ordered from IDT
Add-on orders from IDT
Collaboration with Vincent Lim from National Yang-Ming University> - August 27th
- PCR JAK5' and JAK3'
- August 28th
- PCR JAK5' and JAK3'
- August 29th
- PCR JAK5' and JAK3'
- August 30th
- 1. Restriction digests and ligates JAK5' and JAK3'
2. PCR and gel purification of STAT1
3. PCR and gel purification of HlyD, HlyB and IL-12 - August 31st
- PCR of ligated JAK5'+3'(extension time 3:45)
- SEPTEMBER
- September 1st
- PCR ligated JAK5' and 3' (extension time 3:45)
diluted 100x, 1000x, 10000x restriction digests and gel purificated STAT1.
restriction digests and gel purification of HlyD, HlyB - September 3rd
- Preculture pSB1C3
HlyB＋D ligation→colony PCR - September 4th
- Minipreparation→electrophoresis
HlyB＋D→electrophoresis - September 5th
- Minipreparation→gel purification restriction enzyme treatment (EcoRI, SpeI)
restriction digests PCR-ed HlyB＋D (EcoRI, XbaI)→gel purification - September 6th
- pSB1C3 and HlyD+HlyB ligation⇨transformation
PCR:
1. GFP+HlyA
2. HlyA+GFP
3. HlyA+IL-12
4. IL-12+HlyA
5. diluted TolC - September 7th
- Gel purification of pSB1C3
PCR:
1. GFP+HlyA
2. HlyA+GFP
3. HlyA+IL-12
4. IL-12+HlyA
5. TolC
→Gel purification
Contact: kaitjapan@gmail.com Twitter:@KAIT_JAPAN
