Team:Warsaw/Calendar-Main/23 September 2009

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Gradient PCR PhoP/PhoQ

Kama


Tasks:

  • Amplification of phoP/phoQ

Methods:

  • PCR mixture's composition:

    1μl Pfu buffer (EURX) 
    0,25μl primer phoPF
    0,25μl primer phoQR 
    0,5μl dNTPs (10 mM)
    0,25μl Pfu turbo polymerase (EURX)
    0,5μl template DNA from Salmonella (unknown strain)
    0,75μl DMSO
    The solution was topped up with H2O to 10μl.
  • PCR programs:
  • pho

    4min 95°C 
    (30s 95°C, 1min 55-65°C, 5min 10s 72°C)x35

    10min 72°C
    ~ 7°C
  • Electrophoretic separation on 1% agarose gel

Results:

  • Gel (from left)
  1. control -
  2. 2-4 - samples (annealing temperature increases to from the left to the right)
  3. M - GeneRuler DNA Ladder Mix #SM0333 (Fermentas)

Notes:





April
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
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