IGEM:Missouri University of Science and Technology/2009/Notebook/Cyanobacteria Mediated Filtration of Coal Flue/2014/08/31: Difference between revisions

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(Autocreate 2014/08/31 Entry for IGEM:Missouri_University_of_Science_and_Technology/2009/Notebook/Cyanobacteria_Mediated_Filtration_of_Coal_Flue)
 
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== <b>Date: MM/DD/YY</b> <b>People in lab: Yourname</b> ==
==<b>Date: 8/31/14</b> <b>People in lab: Kira Buckowing</b>==
 
<b><span style="font-size:16px;">Title:</span></b> PCR of GP1 and PAO1 Temp and Gel to check products
<b><span style="font-size:16px;">Title:</span></b> Fill in the fields on this page to create a new notebook entry.
<b>Start Time:</b> 10:45 am
 
<b>Purpose:</b> To get a positive result to nosZ
<b>Start Time:</b>
<b>Protocol:</b> LTM Ed. 2 PCR and Gels
 
<b>Exceptions:</b> PCR 1 and 3  = 2 uL DNA, 2.5 uL primers; PCR 2 and 4 = 7.5 uL DNA, 2.5 uL primers
<b>Purpose:</b>  
 
<b>Protocol:</b> <b>Exceptions:</b>  
 
<b>Products:</b>  
<b>Products:</b>  
{| border="1" cellpadding="5"
{| border= " 1 " cellpadding = " 5 "  
|-
|-
! Sample Label !! Description !! Source Label !! Quantity
! Well !! 1 !! 2 !! 3 !! 4 !! 5 !! 6 !! 7 !! 8 !! 9
|-
|-
| Entry || Entry || Entry || #
| - || Ladder || - || PCR 1 || - || PCR2 || - || PCR 3 || - || PCR4 ||
|-
| Entry || Entry || Entry || #
|}
|}
<b>Products:</b>
<b>Results:</b>
<b>Notes:</b> THe band looks to be the wrong size, but its the first band that has ever shown up for something with nosZ
<b>Stop Time:</b> 3:45 pm
<b>Next:</b> Troubleshooting - Test primers again with controls


{| border="1" cellpadding="5"
==<b>Date: 8/31/14</b> <b>People in lab: Kira Buckowing</b>==
<b><span style="font-size:16px;">Title:</span></b> innoculation of DH5 Alpha stock
<b>Start Time:</b> 4 pm
<b>Purpose:</b> Prep for making comp cells
<b>Protocol:</b> LTM Ed. 2 Innoculation
<b>Exceptions:</b>
<b>Products:</b>
{| border= " 1 " cellpadding = " 5 "  
|-
|-
! Well !! 1 !! 2 !! 3 !! 4 !! 5 !! 6 !! 7 !! 8
! Sample Label !! Description !! Source Label !! Quantity
|-
|-
| Sample || || ||  ||  ||  ||  ||  ||  
| DH5 alphs stock 8/31 || DH5 alpha cells in LB broth || --- || 1
|}
 
 
 
 
 


<b>Results:</b>


<b>Notes:</b>


<b>Stop Time:</b>  
|}
<b>Results:</b>
<b>Notes:</b> Stored in fridge after incubation period
<b>Stop Time:</b> 4:05 pm
<b>Next:</b> Competent cell creation


<b>Next:</b>
==<b>Date: 8/31/14</b> <b>People in lab: Kira Buckowing</b>==
<b><span style="font-size:16px;">Title:</span></b> PCR and Gel of GP1 and PAO1 Temp with controls
<b>Start Time:</b> 4:30 pm
<b>Purpose:</b> Testing primers
<b>Protocol:</b> LTM Ed. 2 PCR and Gels
<b>Exceptions:</b> 1 and 2 - 2 uL DNA with old primers; 3 and 4 = 7.5 uL DNA with old primers; 5 and 6 = 2 uL DNA with new primers; 7 and 8 = 7.5 uL with new primers
<b>Products:</b>
{| border= " 1 " cellpadding = " 5 "
|-
! Well !! 1 !! 2 !! 3 !! 4 !! 5 !! 6 !! 7 !! 8 !! 9 !! 10
|-
| - || Ladder || PCR1 || PCR2 || PCR3 || PCR4 || PCR5 || PCR6 || PCR7 || PCR8 || Ladder ||
|}
<b>Products:</b>
<b>Results:</b> Bands are still too short, by about 1000 base pairs
<b>Notes:</b> Annealing temp or primers may be the issue
<b>Stop Time:</b> 9 pm
<b>Next:</b> More troubleshooting




Revision as of 14:36, 14 October 2014

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Date: 8/31/14 People in lab: Kira Buckowing

Title: PCR of GP1 and PAO1 Temp and Gel to check products Start Time: 10:45 am Purpose: To get a positive result to nosZ Protocol: LTM Ed. 2 PCR and Gels Exceptions: PCR 1 and 3 = 2 uL DNA, 2.5 uL primers; PCR 2 and 4 = 7.5 uL DNA, 2.5 uL primers Products:

Well 1 2 3 4 5 6 7 8 9
- Ladder - PCR 1 - PCR2 - PCR 3 - PCR4

Products: Results: Notes: THe band looks to be the wrong size, but its the first band that has ever shown up for something with nosZ Stop Time: 3:45 pm Next: Troubleshooting - Test primers again with controls

Date: 8/31/14 People in lab: Kira Buckowing

Title: innoculation of DH5 Alpha stock Start Time: 4 pm Purpose: Prep for making comp cells Protocol: LTM Ed. 2 Innoculation Exceptions: Products:

Sample Label Description Source Label Quantity
DH5 alphs stock 8/31 DH5 alpha cells in LB broth --- 1





Results: Notes: Stored in fridge after incubation period Stop Time: 4:05 pm Next: Competent cell creation

Date: 8/31/14 People in lab: Kira Buckowing

Title: PCR and Gel of GP1 and PAO1 Temp with controls Start Time: 4:30 pm Purpose: Testing primers Protocol: LTM Ed. 2 PCR and Gels Exceptions: 1 and 2 - 2 uL DNA with old primers; 3 and 4 = 7.5 uL DNA with old primers; 5 and 6 = 2 uL DNA with new primers; 7 and 8 = 7.5 uL with new primers Products:

Well 1 2 3 4 5 6 7 8 9 10
- Ladder PCR1 PCR2 PCR3 PCR4 PCR5 PCR6 PCR7 PCR8 Ladder

Products: Results: Bands are still too short, by about 1000 base pairs Notes: Annealing temp or primers may be the issue Stop Time: 9 pm Next: More troubleshooting



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