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NOTEBOOK - MAY
- Preparation of Competent Cells, aliquot the competent cells, stored in liquid nitrogen and then -80C refrigerator.
- Transformation of: P123H, P53K, P421E
- Mix content for N.P./ H.S. . Without the repaired casamino acid. RE analysis using XbaI and PstI, there are no expected bands were observed for CadC and there are expected bands were observed for pSBIC
- Mixing cultures for HS and NP
- Make LB agar solution
- Calibrate pH valve for N.P. and H.S. special medium
- Preparation of medium for Natronomonas Pharaonis (N.P.) and Halobacterium Salinarum (H.S.). After autoclave, precipitate formed.
- Re-mix the culture for N.P. and H.S. without autoclaving it. There was no precipitate formation and we successfully made the medium.
- Mixing culture for H.S and N.P as the previous recipe calibration of pH
- Checking of cultured N.P. and H.S. are viable or not by light microscope and there are no cells are found.
- Make plate with antibiotics –Ampicillin.
- Primer dilution of the 20 light ideas primers and sub-culturing of N.P. and H.S..
- Amplification by using PCR, following by run gel of Sensory Rhodopsins Protein II gene.
- Amplification by using PCR, following by run gel of Sensory Rhodopsins Protein I gene.
- Amplification by using PCR, following by run gel of Htr I. Incorrect banding shown that the amplification using PCR fail.
- Make plate with antibiotics-chloramphenicol.
- Amplification by using PCR, following by run gel of Htr II. Correct banding shown and HtrI is being successfully amplified.
- Amplification by using PCR, following by run gel of Tsr I. Correct banding shown and HtrI is being successfully amplified.
- Amplification by using PCR, following by run gel of Tsr II. Incorrect banding shown that the amplification using PCR fail.
- Redo PCR amplification of Htr I, following by run gel. Correct banding shown and HtrI is being successfully amplified.
- Amplification by using PCR, following by run gel of Backbone Vector A. Incorrect banding shown.
- Redo PCR amplification of Tsr II, following by run gel. Correct banding shown and HtrI is being successfully amplified.
- Amplification by using PCR, following by run gel of Backbone Vector B. Incorrect banding shown that the PCR amplification was failed.
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