IGEM:Missouri University of Science and Technology/2009/Notebook/Cyanobacteria Mediated Filtration of Coal Flue/2013/08/06: Difference between revisions
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<b>Purpose:</b> Dilute primers for PCR reaction of norV gene, PCR norV gene from E. coli K-12 add iGEM prefix-suffix | <b>Purpose:</b> Dilute primers for PCR reaction of norV gene, PCR norV gene from E. coli K-12 add iGEM prefix-suffix | ||
<b>Protocol:</b> Dilution: Forward Primer: 0.39mg, To make 1 ug/uL: add 390 uL milliQ, To make 10ng/uL: add 1 uL of 1 ug/uL solution to 100 uL milliQ, Final Solution: ~ | <b>Protocol:</b> Dilution: Forward Primer: 0.39mg, To make 1 ug/uL: add 390 uL milliQ, To make 10ng/uL: add 1 uL of 1 ug/uL solution to 100 uL milliQ, Final Solution: ~??.?? uM | ||
Reverse Primer: 0.38mg, To make 1ug/uL: add 380 uL milliQ, To make 10ng/uL: add 1 uL of 1 ug/uL solution to 100 uL milliQ, Final Solution: ~ | Reverse Primer: 0.38mg, To make 1ug/uL: add 380 uL milliQ, To make 10ng/uL: add 1 uL of 1 ug/uL solution to 100 uL milliQ, Final Solution: ~??.?? uM | ||
<b>Products:</b> | <b>Products:</b> | ||
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<b>Purpose:</b> To PCR amplify nosZ and add iGEM prefix and suffix | <b>Purpose:</b> To PCR amplify nosZ and add iGEM prefix and suffix | ||
<b>Protocol:</b> LTM ed. 2 pg 48 <b>Exceptions:</b> 1. Final primer concentration | <b>Protocol:</b> LTM ed. 2 pg 48 <b>Exceptions:</b> 1. Final primer concentration <0.1 uM | ||
PCR run conditions: Annealing Temp: 65C, elongation time: 5min, template P. aeru 7/30/13 | PCR run conditions: Annealing Temp: 65C, elongation time: 5min, template P. aeru 7/30/13 | ||
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<b>Next:</b> Gel electrophoresis and sequencing | <b>Next:</b> Gel electrophoresis and sequencing | ||
== <b>Date: 08/06/13</b> <b> Emily Puleo, Alie Abele</b> == | |||
<b><span style="font-size:16px;">Title:</span></b> PCR hmp gene from E. coli to add iGEM prefix & suffix | |||
<b>Start Time:</b> 8:10 PM | |||
<b>Purpose:</b> To PCR amplify hmp and add iGEM prefix and suffix | |||
<b>Protocol:</b> LTM ed. 2 pg 48 <b>Exceptions:</b> 1. Final primer concentration <0.1 uM | |||
PCR run conditions: Annealing Temp: 65C, elongation time: 5min, template washed and re-suspended E. coli 7/30/13 | |||
<b>Products:</b> | |||
{| border="1" cellpadding="5" | |||
|- | |||
! Sample Label !! Description !! Source Label !! Quantity | |||
|- | |||
| 8/6/13 A hmp || hmp gene with iGEM prefix&suffix using 7.5 uL template in 0.5 mL H2O || E. coli 7/30/13 || 2 | |||
|- | |||
| 8/6/13 F primer hmp || 10 ng/uL hmp forward primer || hmp Forward Primer || 1 | |||
|- | |||
| 8/6/13 R primer nosZ || 10 ng/uL hmp reverse primer || hmp Reverse Primer || 1 | |||
|} | |||
<b>Results:</b> N/A | |||
<b>Notes:</b> 0.39 mg of F primer + 390 uL of milliQ, diluted 1:100 with milliQ = 10 ng/uL F primer, repeated with 0.36 mg R primer and 360 uL milliQ. NOTE: Corrected math when entered into online notebook, recheck to confirm numbers | |||
<b>Stop Time:</b> 9:00 PM | |||
<b>Next:</b> Gel electrophoresis and sequencing | |||
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Revision as of 12:23, 25 February 2014
 Cyanobacteria Mediated Filtration of Coal Flue Gas
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Date: 08/06/13 Blythe Ferriere, Annie GooTitle: People in lab: Dilute primers plus PCR Start Time: 2:15 PM Purpose: Dilute primers for PCR reaction of norV gene, PCR norV gene from E. coli K-12 add iGEM prefix-suffix Protocol: Dilution: Forward Primer: 0.39mg, To make 1 ug/uL: add 390 uL milliQ, To make 10ng/uL: add 1 uL of 1 ug/uL solution to 100 uL milliQ, Final Solution: ~??.?? uM Reverse Primer: 0.38mg, To make 1ug/uL: add 380 uL milliQ, To make 10ng/uL: add 1 uL of 1 ug/uL solution to 100 uL milliQ, Final Solution: ~??.?? uM Products:
Results: N/A Notes: Changed programs about 15 minutes in Stop Time: 5:00 PM Next: Ligtate into iGEM backbone Date: 08/06/13 Emily Puleo, Alie AbeleTitle: Digestion of 8/3/13 GE norCB with EcoRI and PstI Start Time: 6:17 PM Purpose: Prep norCB for ligation with iGEM standard plasmid. Protocol: LTM ed. 2 pg 37-39. Exceptions: 1. Master Mix: MilliQ - 39uL, EcoRI - 0.3uL, PstI - 0.3uL, 10x Tango buffer - 6uL, use 5uL DNA per reaction Products:
Results: N/A Notes: Stop Time: 9:00 PM Next: Digestion of plasmids and ligation Date: 08/06/13 Emily Puleo, Alie AbeleTitle: PCR nosZ gene from P. aeruginosa to add iGEM prefix & suffix Start Time: 7:32 PM Purpose: To PCR amplify nosZ and add iGEM prefix and suffix Protocol: LTM ed. 2 pg 48 Exceptions: 1. Final primer concentration <0.1 uM PCR run conditions: Annealing Temp: 65C, elongation time: 5min, template P. aeru 7/30/13 Products:
Results: N/A Notes: See 7/31/13 notes for calculations Stop Time: 9:00 PM Next: Gel electrophoresis and sequencing Date: 08/06/13 Emily Puleo, Alie AbeleTitle: PCR hmp gene from E. coli to add iGEM prefix & suffix Start Time: 8:10 PM Purpose: To PCR amplify hmp and add iGEM prefix and suffix Protocol: LTM ed. 2 pg 48 Exceptions: 1. Final primer concentration <0.1 uM PCR run conditions: Annealing Temp: 65C, elongation time: 5min, template washed and re-suspended E. coli 7/30/13 Products:
Results: N/A Notes: 0.39 mg of F primer + 390 uL of milliQ, diluted 1:100 with milliQ = 10 ng/uL F primer, repeated with 0.36 mg R primer and 360 uL milliQ. NOTE: Corrected math when entered into online notebook, recheck to confirm numbers Stop Time: 9:00 PM Next: Gel electrophoresis and sequencing | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||
