Team:Hong Kong-CUHK/DOC NBK JUN

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NOTEBOOK - JUNE

1. Redo amplification by using PCR, following by run gel of Backbone Vector A.
2. Amplification by using PCR, following by run gel of Backbone Vector C. Incorrect banding shown that the PCR amplification was failed.
3. Amplification by using PCR, following by run gel of Backbone Vector D. Incorrect banding shown that the PCR amplification was failed.
4. Redo PCR amplification of Backbone Vector B, following by run gel. Correct banding shown and Backbone Vector B I is being successfully amplified.
5. Redo PCR amplification of Backbone Vector D, following by run gel. Correct banding shown and Backbone Vector D I is being successfully amplified.


6. Overlapping PCR of NpSRII-NpHtrII-Ectsr-backbone which the plasmid is responsible in repelling from blue light.
7. Redo PCR amplification of Backbone Vector C, following by run gel. Correct banding shown and Backbone Vector B I is being successfully amplified.
8. Overlapping PCR of NpSRII-NpHtrII-Ectar-backbone which the plasmid is responsible in attracting by blue light.
9. Overlapping PCR of HsSRI-HsHtrI-Ectsr-backbone which the plasmid is responsible in repelling from orange light.


10. Overlapping PCR of HsSRI-HsHtrI-Ectar-backbone which the plasmid should be responsible in attracting by orange light.
11. Direct transformation of NpSRII-NpHtrII-Ectsr-backbone gene, follow by spread plate.


12. Make plate with 0.5% soft agar with antibiotics- Ampicillin.
13. Direct transformation of NpSRII-NpHtrII-Ectar-backbone gene, follow by spread plate.
14. Direct transformation of HsSRI-HsHtrI-Ectsr-backbone gene, follow by spread plate.
15. Direct transformation of HsSRI-HsHtrI-Ectar-backbone gene, follow by spread plate.


16. Pick clones from transformed NpSRII-NpHtrII-Ectsr-backbone cells.
17. Pick clones from transformed NpSRII-NpHtrII-Ectar-backbone cells. There is no clone on the plate. Direct transformation need to re-do.
18. Pick clones from transformed HsSRI-HsHtrI-Ectsr-backbone cells.


19. Pick clones from transformed HsSRI-HsHtrI-Ectar-backbone cells.
20. Direct transformation of NpSRII-NpHtrII-Ectar-backbone gene, follow by spread plate.
21. Pick clones from transformed NpSRII-NpHtrII-Ectar-backbone cells.
22. Miniprep of picked clones from transformed NpSRII-NpHtrII-Ectsr-backbone cells.


23. Plasmids containing NpSRII-NpHtrII-Ectsr-backbone gene were sent to sequencing.
24. Sequencing results shown that the sequence of plasmids containing NpSRII-NpHtrII-Ectsr-backbone gene were incorrect.

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