Team:Calgary/21 July 2010

From 2010.igem.org

(Difference between revisions)
Line 5: Line 5:
Objective: PCR out native MAlE/ MalE 31 protein out of E.coli
Objective: PCR out native MAlE/ MalE 31 protein out of E.coli
-
Purpose: Testing Mal E/ Mal E31 primers to get 1188 bp long gene of interest. Primers that are used are gene specific and anneals right on the start of the Mal E gene
+
Purpose: Testing Mal E/ Mal E31 primers to get 1188 bp long gene of interest. Primers that are used are gene specific and anneals right on the start of the Mal E gene.
-
'''Method: Preparing the Master Mix'''  
+
'''Tube Labels'''
-
{| class="wikitable"
+
*1- p1H  - C1
-
|-
+
*2- p1H  - C1
-
! Reagent
+
*3- p1H  - C2
-
! x1μL
+
*4- p1H  - C2
-
! x8μL
+
*5- p31H  - C1
-
|-
+
*6- p31H  - C2
-
| 10x PCR Mg<sup>2+</sup>
+
*7- p1ΔSS - C1
-
| 5.0
+
*8- Master Mix
-
| 40.0
+
-
|-
+
-
| 2 mm dNTP
+
-
| 5.0
+
-
| 40.0
+
-
|-
+
-
| 50 mm MgCl<sub>2</sub>
+
-
| 1.5
+
-
| 12.0
+
-
|-
+
-
| ddH<sub>2</sub>O
+
-
| 36.3
+
-
| 290.4
+
-
|-  
+
-
| MalE-N-F 10 mm
+
-
| 1.0
+
-
| 8.0
+
-
|-
+
-
| MalE-N-R 10 mm
+
-
| 1.0
+
-
| 8.0
+
-
|-
+
-
| Platinum Taq Polymerase
+
-
| 0.2
+
-
| 1.6
+
-
|-
+
-
|}
+
'''PCR Conditions'''
'''PCR Conditions'''
Line 50: Line 23:
*Stage 2:52°C  for  0:45 min
*Stage 2:52°C  for  0:45 min
*Stage 3:72°C  for  1:15 min
*Stage 3:72°C  for  1:15 min
-
Cycle 3: 72°C  for 10:00 min
+
*Cycle 3: 72°C  for 10:00 min
Holding Temp: 4°C
Holding Temp: 4°C
}}
}}

Revision as of 22:00, 21 July 2010

Notebook

May
MTWTFSS
          [http://2010.igem.org/Team:Calgary/1_May_2010 1] [http://2010.igem.org/Team:Calgary/2_May_2010 2]
[http://2010.igem.org/Team:Calgary/3_May_2010 3] [http://2010.igem.org/Team:Calgary/4_May_2010 4] [http://2010.igem.org/Team:Calgary/5_May_2010 5] [http://2010.igem.org/Team:Calgary/6_May_2010 6] [http://2010.igem.org/Team:Calgary/7_May_2010 7] [http://2010.igem.org/Team:Calgary/8_May_2010 8] [http://2010.igem.org/Team:Calgary/9_May_2010 9]
[http://2010.igem.org/Team:Calgary/10_May_2010 10] [http://2010.igem.org/Team:Calgary/11_May_2010 11] [http://2010.igem.org/Team:Calgary/12_May_2010 12] [http://2010.igem.org/Team:Calgary/13_May_2010 13] [http://2010.igem.org/Team:Calgary/14_May_2010 14] [http://2010.igem.org/Team:Calgary/15_May_2010 15] [http://2010.igem.org/Team:Calgary/16_May_2010 16]
[http://2010.igem.org/Team:Calgary/17_May_2010 17] [http://2010.igem.org/Team:Calgary/18_May_2010 18] [http://2010.igem.org/Team:Calgary/19_May_2010 19] [http://2010.igem.org/Team:Calgary/20_May_2010 20] [http://2010.igem.org/Team:Calgary/21_May_2010 21] [http://2010.igem.org/Team:Calgary/22_May_2010 22] [http://2010.igem.org/Team:Calgary/23_May_2010 23]
[http://2010.igem.org/Team:Calgary/24_May_2010 24] [http://2010.igem.org/Team:Calgary/25_May_2010 25] [http://2010.igem.org/Team:Calgary/26_May_2010 26] [http://2010.igem.org/Team:Calgary/27_May_2010 27] [http://2010.igem.org/Team:Calgary/28_May_2010 28] [http://2010.igem.org/Team:Calgary/29_May_2010 29] [http://2010.igem.org/Team:Calgary/30_May_2010 30]
[http://2010.igem.org/Team:Calgary/31_May_2010 31]

June
MTWTFSS
  [http://2010.igem.org/Team:Calgary/1_June_2010 1] [http://2010.igem.org/Team:Calgary/2_June_2010 2] [http://2010.igem.org/Team:Calgary/3_June_2010 3] [http://2010.igem.org/Team:Calgary/4_June_2010 4] [http://2010.igem.org/Team:Calgary/5_June_2010 5] [http://2010.igem.org/Team:Calgary/6_June_2010 6]
[http://2010.igem.org/Team:Calgary/7_June_2010 7] [http://2010.igem.org/Team:Calgary/8_June_2010 8] [http://2010.igem.org/Team:Calgary/9_June_2010 9] [http://2010.igem.org/Team:Calgary/10_June_2010 10] [http://2010.igem.org/Team:Calgary/11_June_2010 11] [http://2010.igem.org/Team:Calgary/12_June_2010 12] [http://2010.igem.org/Team:Calgary/13_June_2010 13]
[http://2010.igem.org/Team:Calgary/14_June_2010 14] [http://2010.igem.org/Team:Calgary/15_June_2010 15] [http://2010.igem.org/Team:Calgary/16_June_2010 16] [http://2010.igem.org/Team:Calgary/17_June_2010 17] [http://2010.igem.org/Team:Calgary/18_June_2010 18] [http://2010.igem.org/Team:Calgary/19_June_2010 19] [http://2010.igem.org/Team:Calgary/20_June_2010 20]
[http://2010.igem.org/Team:Calgary/21_June_2010 21] [http://2010.igem.org/Team:Calgary/22_June_2010 22] [http://2010.igem.org/Team:Calgary/23_June_2010 23] [http://2010.igem.org/Team:Calgary/24_June_2010 24] [http://2010.igem.org/Team:Calgary/25_June_2010 25] [http://2010.igem.org/Team:Calgary/26_June_2010 26] [http://2010.igem.org/Team:Calgary/27_June_2010 27]
[http://2010.igem.org/Team:Calgary/28_June_2010 28] [http://2010.igem.org/Team:Calgary/29_June_2010 29] [http://2010.igem.org/Team:Calgary/30_June_2010 30]

July
MTWTFSS
      [http://2010.igem.org/Team:Calgary/1_July_2010 1] [http://2010.igem.org/Team:Calgary/2_July_2010 2] [http://2010.igem.org/Team:Calgary/3_July_2010 3] [http://2010.igem.org/Team:Calgary/4_July_2010 4]
[http://2010.igem.org/Team:Calgary/5_July_2010 5] [http://2010.igem.org/Team:Calgary/6_July_2010 6] [http://2010.igem.org/Team:Calgary/7_July_2010 7] [http://2010.igem.org/Team:Calgary/8_July_2010 8] [http://2010.igem.org/Team:Calgary/9_July_2010 9] [http://2010.igem.org/Team:Calgary/10_July_2010 10] [http://2010.igem.org/Team:Calgary/11_July_2010 11]
[http://2010.igem.org/Team:Calgary/12_July_2010 12] [http://2010.igem.org/Team:Calgary/13_July_2010 13] [http://2010.igem.org/Team:Calgary/14_July_2010 14] [http://2010.igem.org/Team:Calgary/15_July_2010 15] [http://2010.igem.org/Team:Calgary/16_July_2010 16] [http://2010.igem.org/Team:Calgary/17_July_2010 17] [http://2010.igem.org/Team:Calgary/18_July_2010 18]
[http://2010.igem.org/Team:Calgary/19_July_2010 19] [http://2010.igem.org/Team:Calgary/20_July_2010 20] [http://2010.igem.org/Team:Calgary/21_July_2010 21] [http://2010.igem.org/Team:Calgary/22_July_2010 22] [http://2010.igem.org/Team:Calgary/23_July_2010 23] [http://2010.igem.org/Team:Calgary/24_July_2010 24] [http://2010.igem.org/Team:Calgary/25_July_2010 25]
[http://2010.igem.org/Team:Calgary/26_July_2010 26] [http://2010.igem.org/Team:Calgary/27_July_2010 27] [http://2010.igem.org/Team:Calgary/28_July_2010 28] [http://2010.igem.org/Team:Calgary/29_July_2010 29] [http://2010.igem.org/Team:Calgary/30_July_2010 30] [http://2010.igem.org/Team:Calgary/31_July_2010 31]

August
MTWTFSS
            [http://2010.igem.org/Team:Calgary/1_August_2010 1]
[http://2010.igem.org/Team:Calgary/2_August_2010 2] [http://2010.igem.org/Team:Calgary/3_August_2010 3] [http://2010.igem.org/Team:Calgary/4_August_2010 4] [http://2010.igem.org/Team:Calgary/5_August_2010 5] [http://2010.igem.org/Team:Calgary/6_August_2010 6] [http://2010.igem.org/Team:Calgary/7_August_2010 7] [http://2010.igem.org/Team:Calgary/8_August_2010 8]
[http://2010.igem.org/Team:Calgary/9_August_2010 9] [http://2010.igem.org/Team:Calgary/10_August_2010 10] [http://2010.igem.org/Team:Calgary/11_August_2010 11] [http://2010.igem.org/Team:Calgary/12_August_2010 12] [http://2010.igem.org/Team:Calgary/13_August_2010 13] [http://2010.igem.org/Team:Calgary/14_August_2010 14] [http://2010.igem.org/Team:Calgary/15_August_2010 15]
[http://2010.igem.org/Team:Calgary/16_August_2010 16] [http://2010.igem.org/Team:Calgary/17_August_2010 17] [http://2010.igem.org/Team:Calgary/18_August_2010 18] [http://2010.igem.org/Team:Calgary/19_August_2010 19] [http://2010.igem.org/Team:Calgary/20_August_2010 20] [http://2010.igem.org/Team:Calgary/21_August_2010 21] [http://2010.igem.org/Team:Calgary/22_August_2010 22]
[http://2010.igem.org/Team:Calgary/23_August_2010 23] [http://2010.igem.org/Team:Calgary/24_August_2010 24] [http://2010.igem.org/Team:Calgary/25_August_2010 25] [http://2010.igem.org/Team:Calgary/26_August_2010 26] [http://2010.igem.org/Team:Calgary/27_August_2010 27] [http://2010.igem.org/Team:Calgary/28_August_2010 28] [http://2010.igem.org/Team:Calgary/29_August_2010 29]
[http://2010.igem.org/Team:Calgary/30_August_2010 30] [http://2010.igem.org/Team:Calgary/31_August_2010 31]

September
MTWTFSS
    [http://2010.igem.org/Team:Calgary/1_September_2010 1] [http://2010.igem.org/Team:Calgary/2_September_2010 2] [http://2010.igem.org/Team:Calgary/3_September_2010 3] [http://2010.igem.org/Team:Calgary/4_September_2010 4] [http://2010.igem.org/Team:Calgary/5_September_2010 5]
[http://2010.igem.org/Team:Calgary/6_September_2010 6] [http://2010.igem.org/Team:Calgary/7_September_2010 7] [http://2010.igem.org/Team:Calgary/8_September_2010 8] [http://2010.igem.org/Team:Calgary/9_September_2010 9] [http://2010.igem.org/Team:Calgary/10_September_2010 10] [http://2010.igem.org/Team:Calgary/11_September_2010 11] [http://2010.igem.org/Team:Calgary/12_September_2010 12]
[http://2010.igem.org/Team:Calgary/13_September_2010 13] [http://2010.igem.org/Team:Calgary/14_September_2010 14] [http://2010.igem.org/Team:Calgary/15_September_2010 15] [http://2010.igem.org/Team:Calgary/16_September_2010 16] [http://2010.igem.org/Team:Calgary/17_September_2010 17] [http://2010.igem.org/Team:Calgary/18_September_2010 18] [http://2010.igem.org/Team:Calgary/19_September_2010 19]
[http://2010.igem.org/Team:Calgary/20_September_2010 20] [http://2010.igem.org/Team:Calgary/21_September_2010 21] [http://2010.igem.org/Team:Calgary/22_September_2010 22] [http://2010.igem.org/Team:Calgary/23_September_2010 23] [http://2010.igem.org/Team:Calgary/24_September_2010 24] [http://2010.igem.org/Team:Calgary/25_September_2010 25] [http://2010.igem.org/Team:Calgary/26_September_2010 26]
[http://2010.igem.org/Team:Calgary/27_September_2010 27] [http://2010.igem.org/Team:Calgary/28_September_2010 28] [http://2010.igem.org/Team:Calgary/29_September_2010 29] [http://2010.igem.org/Team:Calgary/30_September_2010 30]

October
MTWTFSS
        [http://2010.igem.org/Team:Calgary/1_October_2010 1] [http://2010.igem.org/Team:Calgary/2_October_2010 2] [http://2010.igem.org/Team:Calgary/3_October_2010 3]
[http://2010.igem.org/Team:Calgary/4_October_2010 4] [http://2010.igem.org/Team:Calgary/5_October_2010 5] [http://2010.igem.org/Team:Calgary/6_October_2010 6] [http://2010.igem.org/Team:Calgary/7_October_2010 7] [http://2010.igem.org/Team:Calgary/8_October_2010 8] [http://2010.igem.org/Team:Calgary/9_October_2010 9] [http://2010.igem.org/Team:Calgary/10_October_2010 10]
[http://2010.igem.org/Team:Calgary/11_October_2010 11] [http://2010.igem.org/Team:Calgary/12_October_2010 12] [http://2010.igem.org/Team:Calgary/13_October_2010 13] [http://2010.igem.org/Team:Calgary/14_October_2010 14] [http://2010.igem.org/Team:Calgary/15_October_2010 15] [http://2010.igem.org/Team:Calgary/16_October_2010 16] [http://2010.igem.org/Team:Calgary/17_October_2010 17]
[http://2010.igem.org/Team:Calgary/18_October_2010 18] [http://2010.igem.org/Team:Calgary/19_October_2010 19] [http://2010.igem.org/Team:Calgary/20_October_2010 20] [http://2010.igem.org/Team:Calgary/21_October_2010 21] [http://2010.igem.org/Team:Calgary/22_October_2010 22] [http://2010.igem.org/Team:Calgary/23_October_2010 23] [http://2010.igem.org/Team:Calgary/24_October_2010 24]
[http://2010.igem.org/Team:Calgary/25_October_2010 25] [http://2010.igem.org/Team:Calgary/26_October_2010 26] [http://2010.igem.org/Team:Calgary/27_October_2010 27] [http://2010.igem.org/Team:Calgary/28_October_2010 28] [http://2010.igem.org/Team:Calgary/29_October_2010 29] [http://2010.igem.org/Team:Calgary/30_October_2010 30] [http://2010.igem.org/Team:Calgary/31_October_2010 31]

Wednesday July 21, 2010

Raida:

Objective: PCR out native MAlE/ MalE 31 protein out of E.coli Purpose: Testing Mal E/ Mal E31 primers to get 1188 bp long gene of interest. Primers that are used are gene specific and anneals right on the start of the Mal E gene.

Tube Labels

  • 1- p1H - C1
  • 2- p1H - C1
  • 3- p1H - C2
  • 4- p1H - C2
  • 5- p31H - C1
  • 6- p31H - C2
  • 7- p1ΔSS - C1
  • 8- Master Mix

PCR Conditions Cycle 1: 94°C 2:00 min Cycle 2:

  • Stage 1:94°C for 1:00 min
  • Stage 2:52°C for 0:45 min
  • Stage 3:72°C for 1:15 min
  • Cycle 3: 72°C for 10:00 min

Holding Temp: 4°C


No notebook page exists for this date. Sorry!

Retrieved from "http://2010.igem.org/Team:Calgary/21_July_2010"