Team:Hong Kong-CUHK/DOC NBK MAY
From 2012.igem.org
(Difference between revisions)
(2 intermediate revisions not shown) | |||
Line 8: | Line 8: | ||
<td style="background-color:#FFF; padding:25px"><!-- InstanceBeginEditable name="EditRegion1" --> | <td style="background-color:#FFF; padding:25px"><!-- InstanceBeginEditable name="EditRegion1" --> | ||
<p> </p> | <p> </p> | ||
- | <p class="aloveofthunder" style="line-height:normal">NOTEBOOK - MAY</p> | + | <p class="aloveofthunder" style="line-height:normal; margin-bottom:35px">NOTEBOOK - MAY</p> |
<ol> | <ol> | ||
- | <li>Preparation of Competent Cells, aliquot the competent cells, stored in liquid | + | <li>Preparation of Competent Cells, aliquot the competent cells, stored in liquid nitrogen and then -80C refrigerator. </li> |
<li>Transformation of: P123H, P53K, P421E</li> | <li>Transformation of: P123H, P53K, P421E</li> | ||
- | + | </br> | |
- | <li>Mix content for N.P./ H.S. . Without the repaired casamino acid. RE analysis | + | <li>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</li> |
<li>Mixing cultures for HS and NP</li> | <li>Mixing cultures for HS and NP</li> | ||
<li>Make LB agar solution</li> | <li>Make LB agar solution</li> | ||
- | + | </br> | |
<li>Calibrate pH valve for N.P. and H.S. special medium</li> | <li>Calibrate pH valve for N.P. and H.S. special medium</li> | ||
- | <li>Preparation of medium for Natronomonas Pharaonis(N.P.) and | + | <li>Preparation of medium for <i>Natronomonas Pharaonis</i> (N.P.) and <i>Halobacterium Salinarum</i> (H.S.). After autoclave, precipitate formed. </li> |
- | <li>Re-mix the culture for N.P. and H.S. without autoclaving it. There was no | + | <li>Re-mix the culture for N.P. and H.S. without autoclaving it. There was no precipitate formation and we successfully made the medium.</li> |
- | + | </br> | |
<li>Mixing culture for H.S and N.P as the previous recipe calibration of pH </li> | <li>Mixing culture for H.S and N.P as the previous recipe calibration of pH </li> | ||
- | <li>Checking of cultured N.P. and H.S. are viable or not by light microscope and | + | <li>Checking of cultured N.P. and H.S. are viable or not by light microscope and there are no cells are found. </li> |
- | <li>Make plate with antibiotics | + | <li>Make plate with antibiotics –Ampicillin. </li> |
<li>Primer dilution of the 20 light ideas primers and sub-culturing of N.P. and H.S.. </li> | <li>Primer dilution of the 20 light ideas primers and sub-culturing of N.P. and H.S.. </li> | ||
<li>Amplification by using PCR, following by run gel of Sensory Rhodopsins Protein II gene. </li> | <li>Amplification by using PCR, following by run gel of Sensory Rhodopsins Protein II gene. </li> | ||
<li>Amplification by using PCR, following by run gel of Sensory Rhodopsins Protein I gene. </li> | <li>Amplification by using PCR, following by run gel of Sensory Rhodopsins Protein I gene. </li> | ||
- | + | </br> | |
- | <li>Amplification by using PCR, following by run gel of Htr I. Incorrect banding | + | <li>Amplification by using PCR, following by run gel of Htr I. Incorrect banding shown that the amplification using PCR fail. </li> |
<li>Make plate with antibiotics-chloramphenicol. </li> | <li>Make plate with antibiotics-chloramphenicol. </li> | ||
- | <li>Amplification by using PCR, following by run gel of Htr II. Correct banding | + | <li>Amplification by using PCR, following by run gel of Htr II. Correct banding shown and HtrI is being successfully amplified. </li> |
- | <li>Amplification by using PCR, following by run gel of Tsr I. Correct banding | + | <li>Amplification by using PCR, following by run gel of Tsr I. Correct banding shown and HtrI is being successfully amplified. </li> |
- | + | </br> | |
- | <li>Amplification by using PCR, following by run gel of Tsr II. Incorrect | + | <li>Amplification by using PCR, following by run gel of Tsr II. Incorrect banding shown that the amplification using PCR fail. </li> |
- | <li>Redo PCR amplification of Htr I, following by run gel. Correct banding | + | <li>Redo PCR amplification of Htr I, following by run gel. Correct banding shown and HtrI is being successfully amplified. </li> |
- | <li>Amplification by using PCR, following by run gel of Backbone Vector A. | + | <li>Amplification by using PCR, following by run gel of Backbone Vector A. Incorrect banding shown. </li> |
- | + | </br> | |
- | <li>Redo PCR amplification of Tsr II, following by run gel. Correct banding | + | <li>Redo PCR amplification of Tsr II, following by run gel. Correct banding shown and HtrI is being successfully amplified. </li> |
- | <li>Amplification by using PCR, following by run gel of Backbone Vector B. | + | <li>Amplification by using PCR, following by run gel of Backbone Vector B. Incorrect banding shown that the PCR amplification was failed. </li> |
- | + | ||
</ol> | </ol> | ||
<p> </p> | <p> </p> |
Latest revision as of 02:23, 27 September 2012
![]() |
Check out our FACEBOOK page! |
NOTEBOOK - MAY
|
Home | Team | Project | Biobricks | Human Practice | Safety | Documentation | Acknowledgement
Address: Rm. 184, Science Centre, CUHK Copyright © 2012 Apycom jQuery Menus |