October
4/10/2010
- digest of 8 fragments with BsaI-HF @ 37°C for 1hr
- 6x2ul of F1-8
- 3ul NEB4 10x
- 3ul BSA 10x
- 11ul H2O
- 1ul BsaI-HF
- purification with Nucleotide Removal Kit
- NanoDrop concentration check:
| Fragment 1
| Fragment 2
| Fragment 3
| Fragment 4
| Fragment 5
| Fragment 6
| Fragment 7
| Fragment 8
|
| 35.5 ng/ul
| 43.5 ng/ul
| 43.0 ng/ul
| 43.0 ng/ul
| 79.0 ng/ul
| 51.0 ng/ul
| 63.0 ng/ul
| 44.0 ng/ul
|
5/10/2010
- assembly started with Quick ligase kit
- ~200ng of fragment DNA per reaction
- ligation time ~10min
27/09/2010
AAV 1 fragments 1-4; 5-8 and 1-8 (full capsid) assembly
- Gel loading scheme:
- M - GeneRuler 1kb DNA ladder
- F 1-4 - fragments 1-4 assembly
- F 5-8 - fragments 5-8 assembly
- F 1-8 - fragments 1-8 assembly
- C 1-4 - fragments 1-4 control
- C 5-8 - fragments 5-8 control
- C 1-8 - fragments 1-8 control
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