Team:Hong Kong-CUHK/DOC NBK AUG

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NOTEBOOK - AUG

1. Miniprep of picked clones from transformed HsSRI-HsHtrI-Ectar-backbone cells.
2. .
3. Make LB agar solution.
4. Sequencing results shown that the sequence of plasmids containing HsSRI-HsHtrI-Ectsr-backbone gene are incorrect. Production of HsSRI-HsHtrI-Ectsr is fail.
5. The sequenced cells which contain the HsSRI-HsHtrI-Ectsr-backbone gene are being cultured in 0.5% soft agar plate for testing using normal light bulb with its corresponding antibiotics.
6.  
7. Repeat in testing cells that contain the HsSRI-HsHtrI-Ectsr-backbone gene are being cultured in 0.5% soft agar plate for testing using normal light bulb with its corresponding antibiotics.
8. Plasmids containing HsSRI-HsHtrI-Ectar -backbone gene were sent to sequencing.
9.  
10. Plasmids containing NpSRII-NpHtrII-Ectsr-backbone gene were sent to sequencing.
11. Sequencing results shown that the sequence of plasmids containing NpSRII-NpHtrII-Ectsr-backbone gene are correct.
12. The sequenced cells which contain the NpSRII-NpHtrII-Ectsr-backbone gene are being cultured in 0.5% soft agar plate for testing using normal light bulb with its corresponding antibiotics.
13.  
14. Repeat in testing cells that contain the NpSRII-NpHtrII-Ectsr-backbone gene are being cultured in 0.5% soft agar plate for testing using normal light bulb with its corresponding antibiotics.
15. Sequencing results shown that the sequence of plasmids containing HsSRI-HsHtrI-Ectar -backbone gene is correct.
16. .
17. The sequenced cells which contain the HsSRI-HsHtrI-Ectar-backbone gene are being cultured in 0.5% soft agar plate for testing using normal light bulb with its corresponding antibiotics.
18. Repeat in testing cells that contain the HsSRI-HsHtrI-Ectar-backbone gene are being cultured in 0.5% soft agar plate for testing using normal                    light bulb with its corresponding antibiotics.
19. Altering the original backbone that contain NpSRII-NpHtrII-Ectsr gene to the standard backbone. Starting with enzyme restriction cut.
20. Run gel of product from enzyme restriction cut (NpSRII-NpHtrII-Ectsr gene) and the size of the bandings are correct, followed by gel                                                purification.
21. .
22. Ligation is done and the product is being transformed to Top10 competent cells, followed by spread plate.
23. Repeat testing for the sequenced cells which contain the HsSRI-HsHtrI-Ectar-backbone gene, cells are being cultured in 0.5% soft agar plate for testing using normal light bulb with its corresponding antibiotics.
24. Altering the original backbone that contain NpSRII-NpHtrII-Ectar gene to the standard backbone. Starting with enzyme restriction cut.
25. Run gel of product from enzyme restriction cut (NpSRII-NpHtrII-Ectar gene) and the size of the bandings are incorrect, therefore enzyme restriction cut was being re-do.
26. Repeat testing for the sequenced cells which contain the HsSRI-HsHtrI-Ectsr-backbone gene, cells are being cultured in 0.5% soft agar plate for testing using normal light bulb with its corresponding antibiotics.
27. Run gel of product from enzyme restriction cut (NpSRII-NpHtrII-Ectar gene) and the size of the bandings are correct, followed by gel purification.
28. Repeat testing for the sequenced cells which contain the NpSRII-NpHtrII-Ectsr-backbone gene, cells are being cultured in 0.5% soft agar plate for testing using normal light bulb with its corresponding antibiotics.
29. Ligation is done and the product is being transformed to Top10 competent cells, followed by spread plate.
30. Altering the original backbone that contain HsSRI-HsHtrI-Ectar gene to the standard backbone. Starting with enzyme restriction cut.

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