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Team:Wageningen UR/Journal/week14
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* The four 'brickable' TuYV CP gene amplicons (1,1H,4,4H) were digested, ligated into the backbone from BBa_J04450, which is AMP resistant and transformed into our electrocompetent Mach1 ''E coli'' Mach 1. | * The four 'brickable' TuYV CP gene amplicons (1,1H,4,4H) were digested, ligated into the backbone from BBa_J04450, which is AMP resistant and transformed into our electrocompetent Mach1 ''E coli'' Mach 1. | ||
* Colony check: there were many colonies on the sample plate. No colonies were on the negative plate. | * Colony check: there were many colonies on the sample plate. No colonies were on the negative plate. | ||
| - | * We did two colonies PCR check for the samples, but we got negative results from the colony PCR, but we continued to miniprep those colonoes which were used for colony PCR and checked them with normal PCR, this time we saw expected bands. | + | * We did two colonies PCR check for the samples, but we got negative results from the colony PCR, but we continued to miniprep those colonoes which were used for colony PCR and checked them with normal PCR later, this time we saw expected bands for all 4 samples, which should be around 600bp and 800bp. |
| + | [[File:PCR check for miniprep products (TuYV in ampicillin resistant backbone).jpg]] | ||
Revision as of 06:33, 19 September 2012
week 14: 30 july - 5 august
Lab work
Hepatitis B
30.July
- Miniprepping of successful transformants from the transformation (Hep B + BBa_J04500)
- Digestion check of those minipreps
1.August
-> the digestion check was repeated 2 times (once with a longer incubation time and once with a higher amount of DNA in the digestion mixture) until the insert size could be seen on the gel
digestion check 1.August
-> the digestion check confirmed the right insert sizes in the samples they where expected (around 800bp)
TuYV
- The four 'brickable' TuYV CP gene amplicons (1,1H,4,4H) were digested, ligated into the backbone from BBa_J04450, which is AMP resistant and transformed into our electrocompetent Mach1 E coli Mach 1.
- Colony check: there were many colonies on the sample plate. No colonies were on the negative plate.
- We did two colonies PCR check for the samples, but we got negative results from the colony PCR, but we continued to miniprep those colonoes which were used for colony PCR and checked them with normal PCR later, this time we saw expected bands for all 4 samples, which should be around 600bp and 800bp.
.jpg)
PLRV
The PCR products obtained after Reverse Transcription followed by PCR were used as template to add iGEM prefix and suffix.
