Team:HokkaidoU Japan/Notebook/September3
From 2010.igem.org
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* 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`t, so there is posibility that 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 07:34, 1 October 2010
Resultsof yesterdays trnsformation
- 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