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'''  
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'''Tube Labels'''
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{| class="wikitable"
+
*1- p1H  - C1
-
|-
+
*2- p1H  - C1
-
! Reagent
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*3- p1H  - C2
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! x1μL
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*4- p1H  - C2
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! x8μL
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*5- p31H  - C1
-
|-
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*6- p31H  - C2
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| 10x PCR Mg<sup>2+</sup>
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*7- p1ΔSS - C1
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| 5.0
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*8- Master Mix
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| 40.0
+
-
|-
+
-
| 2 mm dNTP
+
-
| 5.0
+
-
| 40.0
+
-
|-
+
-
| 50 mm MgCl<sub>2</sub>
+
-
| 1.5
+
-
| 12.0
+
-
|-
+
-
| ddH<sub>2</sub>O
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| 36.3
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| 290.4
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|-  
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| MalE-N-F 10 mm
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| 1.0
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| 8.0
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-
|-
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| MalE-N-R 10 mm
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| 1.0
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| 8.0
+
-
|-
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| Platinum Taq Polymerase
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| 0.2
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| 1.6
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-
|-
+
-
|}
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'''PCR Conditions'''
'''PCR Conditions'''
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*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
          1 2
3 4 5 6 7 8 9
10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 26 27 28 29 30
31

June
MTWTFSS
  1 2 3 4 5 6
7 8 9 10 11 12 13
14 15 16 17 18 19 20
21 22 23 24 25 26 27
28 29 30

July
MTWTFSS
      1 2 3 4
5 6 7 8 9 10 11
12 13 14 15 16 17 18
19 20 21 22 23 24 25
26 27 28 29 30 31

August
MTWTFSS
            1
2 3 4 5 6 7 8
9 10 11 12 13 14 15
16 17 18 19 20 21 22
23 24 25 26 27 28 29
30 31

September
MTWTFSS
    1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30

October
MTWTFSS
        1 2 3
4 5 6 7 8 9 10
11 12 13 14 15 16 17
18 19 20 21 22 23 24
25 26 27 28 29 30 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"