SBB09Ntbk-SamNg: Difference between revisions

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==~~'''Protocols'''~~==
[http://openwetware.org/wiki/EspP_Autotransporter EspP_Autotransporter]<br>
[http://openwetware.org/wiki/EspP_Autotransporter EspP_Autotransporter]<br>
[http://openwetware.org/wiki/Myc_Tag Myc_Tag]<br>
[http://openwetware.org/wiki/Myc_Tag Myc_Tag]<br>
~~ Under Construction ~~
 
==~~*'''[[User:Sam Ng|Sam Ng]] 9 March 2009''':~~==
'''Completed Protocols:''' <br>
Myc_Tag - Analytical Digest <br>
Myc_Tag - Sequencing <br>
 
'''Additional Notes:''' <br>
N/A
 
==~~*'''[[User:Sam Ng|Sam Ng]] 6 March 2009''':~~==
'''Completed Protocols:''' <br>
EspP Autotransporter - Sequencing <br>
Myc_Tag - Picking of colonies <br>
Myc_Tag - Miniprep purification of DNA <br>
 
'''Additional Notes:''' <br>
N/A
 
<pre>
For Monday Proceed With:
- Myc_Tag - Restriction Digest Check
</pre>
 
==~~*'''[[User:Sam Ng|Sam Ng]] 4 March 2009''':~~==
'''Completed Protocols:''' <br>
EspP Autotransporter - Analytical Digest <br>
Myc_Tag - LR Gateway Transfer<br>
Myc_Tag - Transformation by Heat Shock<br>
 
'''Additional Notes:''' <br>
N/A
 
<pre>
For Monday Proceed With:
- EspP Autotransporter - Sequencing
</pre>
 
==~~*'''[[User:Sam Ng|Sam Ng]] 27 February 2009''':~~==
'''Completed Protocols:''' <br>
EspP Autotransporter - Picking of colonies <br>
EspP Autotransporter - Miniprep purification of DNA <br>
 
'''Additional Notes:''' <br>
Went into Restriction Digestion Check, but incorrectly used all 5 ul, immediately stopped reaction and proceeded with Zymo Cleanup. <br>
 
<pre>
For Wednesday Proceed With:
- EspP Autotransporter - Restriction Digest Check (8 ul)
</pre>
 
==~~*'''[[User:Sam Ng|Sam Ng]] 27 February 2009''':~~==
'''Completed Protocols:''' <br>
EspP Autotransporter - Ligation of EcoRI/BamHI digests <br>
EspP Autotransporter - Transformation by heat-shock <br>
 
'''Additional Notes:''' <br>
N/A
 
<pre>
For Monday Proceed With:
- EspP Autotransporter - Picking of colonies
</pre>
 
==~~*'''[[User:Sam Ng|Sam Ng]] 25 February 2009''':~~==
'''Completed Protocols:''' <br>
EspP Autotransporter - EcoRI/BamHI Digest of PCR Products <br>
EspP Autotransporter - Regular Zymo Cleanup (2) <br>
 
'''Additional Notes:''' <br>
Regular Zymo Cleanup (33ul) **Should have been 8ul the volume of digestion**
 
<pre>
For Friday Proceed With:
- EspP Autotransporter - Ligation of EcoRI/BamHI digests
</pre>


==~~*'''[[User:Sam Ng|Sam Ng]] 23 February 2009''':~~==
==~~*'''[[User:Sam Ng|Sam Ng]] 23 February 2009''':~~==
Ran Analytical Gel (33ul)
'''Completed Protocols:''' <br>
EspP Autotransporter - Analytical Gel <br>
EspP Autotransporter - Regular Zymo Cleanup (1) <br>
 
'''Additional Notes:''' <br>
Lane 3 (~1.5kb)
[[http://openwetware.org/wiki/Image:IMG_1608.JPG]]


Zymo Cleanup (33ul)
Regular Zymo Cleanup (33ul)


<pre>
<pre>
For Wednesday:  
For Wednesday Proceed With:
- Digestion
- EspP Autotransporter - EcoRI/BamHI Digest of PCR Products
</pre>
</pre>


==~~*'''[[User:Sam Ng|Sam Ng]] 18 February 2009''':~~==
==~~*'''[[User:Sam Ng|Sam Ng]] 18 February 2009''':~~==
Received oligos Osn0001-F and Osn0002-R:
'''Completed Protocols:''' <br>
EspP Autotransporter - Oligos <br>
EspP Autotransporter - Cloning by PCR <br>


Osn0001 - 27.00 nmol and Osn0002 - 23.60 nmol
'''Additional Notes:''' <br>
Received oligos Osn0001-F and Osn0002-R: <br>
Osn0001 - 27.00 nmol and Osn0002 - 23.60 nmol <br>


Ran PCR with EHEC
Ran PCR with EHEC <br>
<pre>
<pre>
For Monday:  
For Monday Proceed With:  
- Analytical Gel
- EspP Autotransporter - Analytical Gel
- Cleanup
- EspP Autotransporter - Regular Zymo Cleanup (1)
</pre>
</pre>

Latest revision as of 11:53, 9 March 2009

~~Protocols~~

EspP_Autotransporter
Myc_Tag

~~*Sam Ng 9 March 2009:~~

Completed Protocols:
Myc_Tag - Analytical Digest
Myc_Tag - Sequencing

Additional Notes:
N/A

~~*Sam Ng 6 March 2009:~~

Completed Protocols:
EspP Autotransporter - Sequencing
Myc_Tag - Picking of colonies
Myc_Tag - Miniprep purification of DNA

Additional Notes:
N/A

For Monday Proceed With:
- Myc_Tag - Restriction Digest Check

~~*Sam Ng 4 March 2009:~~

Completed Protocols:
EspP Autotransporter - Analytical Digest
Myc_Tag - LR Gateway Transfer
Myc_Tag - Transformation by Heat Shock

Additional Notes:
N/A

For Monday Proceed With:
- EspP Autotransporter - Sequencing

~~*Sam Ng 27 February 2009:~~

Completed Protocols:
EspP Autotransporter - Picking of colonies
EspP Autotransporter - Miniprep purification of DNA

Additional Notes:
Went into Restriction Digestion Check, but incorrectly used all 5 ul, immediately stopped reaction and proceeded with Zymo Cleanup.

For Wednesday Proceed With:
- EspP Autotransporter - Restriction Digest Check (8 ul)

~~*Sam Ng 27 February 2009:~~

Completed Protocols:
EspP Autotransporter - Ligation of EcoRI/BamHI digests
EspP Autotransporter - Transformation by heat-shock

Additional Notes:
N/A

For Monday Proceed With:
- EspP Autotransporter - Picking of colonies

~~*Sam Ng 25 February 2009:~~

Completed Protocols:
EspP Autotransporter - EcoRI/BamHI Digest of PCR Products
EspP Autotransporter - Regular Zymo Cleanup (2)

Additional Notes:
Regular Zymo Cleanup (33ul) **Should have been 8ul the volume of digestion**

For Friday Proceed With:
- EspP Autotransporter - Ligation of EcoRI/BamHI digests

~~*Sam Ng 23 February 2009:~~

Completed Protocols:
EspP Autotransporter - Analytical Gel
EspP Autotransporter - Regular Zymo Cleanup (1)

Additional Notes:
Lane 3 (~1.5kb) [[1]]

Regular Zymo Cleanup (33ul)

For Wednesday Proceed With:
- EspP Autotransporter - EcoRI/BamHI Digest of PCR Products

~~*Sam Ng 18 February 2009:~~

Completed Protocols:
EspP Autotransporter - Oligos
EspP Autotransporter - Cloning by PCR

Additional Notes:
Received oligos Osn0001-F and Osn0002-R:
Osn0001 - 27.00 nmol and Osn0002 - 23.60 nmol

Ran PCR with EHEC

For Monday Proceed With: 
- EspP Autotransporter - Analytical Gel
- EspP Autotransporter - Regular Zymo Cleanup (1)