"
Team:HokkaidoU Japan/Notebook/September3
From 2010.igem.org
(Difference between revisions)
(→PCR of 1-3A) |
(→Resultsof yesterdays trnsformation) |
||
| Line 1: | Line 1: | ||
{{Template:HokkaidoU_Japan}}<div class="linkbar"><div class="prev">[[Team:HokkaidoU_Japan/Notebook/September2|September 2]]</div>[[Team:HokkaidoU_Japan/Notebook|Notebook]]<div class="next">[[Team:HokkaidoU_Japan/Notebook/September6|September 6]]</div></div> | {{Template:HokkaidoU_Japan}}<div class="linkbar"><div class="prev">[[Team:HokkaidoU_Japan/Notebook/September2|September 2]]</div>[[Team:HokkaidoU_Japan/Notebook|Notebook]]<div class="next">[[Team:HokkaidoU_Japan/Notebook/September6|September 6]]</div></div> | ||
| - | =Resultsof yesterdays | + | =Resultsof yesterdays transformation= |
* pSB1C3 uterly failed to produce colonies | * pSB1C3 uterly failed to produce colonies | ||
* pUC119 produced 20 colonies | * pUC119 produced 20 colonies | ||
| - | * colonies that should been red because of RFP insert wasn | + | * colonies that should been red because of RFP insert wasn't, so there is posibility that insert wasn't there |
=Colony PCR on yesterdays E.coli= | =Colony PCR on yesterdays E.coli= | ||
Revision as of 09:15, 2 October 2010
since 13 Jul 2010
since 1 Aug 2010
Resultsof yesterdays transformation
- pSB1C3 uterly failed to produce colonies
- pUC119 produced 20 colonies
- colonies that should been red because of RFP insert wasn't, so there is posibility that insert wasn't there
Colony PCR on yesterdays E.coli
- Colony PCR was done acordig to protocol
- This day we did 20 samples
Electrophoresis
Concentration check of parts used for transformation yesterday
Confirmed that DNA solution concentration of parts for Ligation was as anticipated (good?)
| Lane | DNA |
| 1 | |
| 2 | Lambda/Hind III, EcoR I |
| 3 | |
| 4 | RFP |
| 5 | pUC119 |
| 6 | pSB1C3 |
PCR of 1-3A
Transformation of 1-3A part which is originaly on pSB1C3 was succesful
Thinking that linearized vector might gone bad we PCRed pSB1C3 from 1-3A
| Reagent | Amount |
|---|---|
| 1-3A | 1 |
| DW | 33 |
| 10x Buffer | 5 |
| 2 M 4dNTPs | 5 |
| 25 mM MgSO4 | 3 |
| Suffix-F | 1 |
| Prefix-R | 1 |
| KOD | 1 |
| Total | 50 uL |
- Extension was for120 sec
- Purified via Microcon YM-10
- Final volume was 43 uL
