# User:Allison K. Alix/Notebook/Thesis Research/2013/06/28

(Difference between revisions)
 Revision as of 10:23, 28 June 2013 (view source) (→Entry title)← Previous diff Revision as of 10:43, 28 June 2013 (view source) (→FCS Sample Prep)Next diff → Line 6: Line 6: | colspan="2"| | colspan="2"| - ==FCS Sample Prep== + ==FCS Measurements== * Run the following samples of MB/DNA * Run the following samples of MB/DNA - 1) 5nM MB 4nM DNA + 1) 10nM MB + 8nM DNA - 2) 1nM MB 960pM DNA + 2) 5nM MB 4nM DNA - 3) 500pM MB 400pM DNA + 3) 1nM MB 960pM DNA - 4) 100pM MB 80pM DNA + 4) 500pM MB 400pM DNA - 5) 50pM MB 40pM MB + 5) 100pM MB 80pM DNA - 6) 25pM MB 20pM DNA + 6) 50pM MB 40pM MB + + 7) 25pM MB 20pM DNA * Run the following oligio-D samples * Run the following oligio-D samples Line 33: Line 35: 5) 100pM 5) 100pM + + ==Calculations== + + MOLECULAR BEACON: + + 7.61μM (x) = 20nM (1000uL) + + x = 2.62uL 7.61uM MB in 997.38uL PBS + + 20nM (x) = (30uL)(10nM) + + 15uL in 15uL PBS + + 20nM (x) = 2nM(100uL) + + 10uL in 90uL + + 2nM (x) = 1nM (100uL) + + x = 50uL in 50uL PBS + + 200pM (100uL) = x (1nM) + + x = 20uL in 80uL PBS + + 100pM(100uL) = (200pM)(x) + + x = 50uL in 50uL buffer + + 50pM (30uL) = 100pM (x) + + x = 15uL in 15uL + + DNA: + + 5.31uM (500uL) = x (53.1uM) + + x = 50uL in 450uL PBS + + 5.31uM (x) = 16nM (1000uL) + + x = 3uL in 997uL PBS + + 16nM (x) = (8nM) (100uL) + + x = 50uL in 50uL PBS + + 1.92nM (100uL) = x (8nM) + + x = 24uL in 76 uL PBS + + 0.8nM (100uL) = x (1.92nM) + + x = 41.7uL in 58.3uL + + 160pM (100) = x (800pM) + + x = 20uL in 80uL + + 80pM (100uL) = x (160pM) + + x = 50uL in 50uL + + 40pM (30uL) = x (80pm) + + x = 15uL in 15uL PBS + + +

## Revision as of 10:43, 28 June 2013

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## FCS Measurements

• Run the following samples of MB/DNA

1) 10nM MB + 8nM DNA

2) 5nM MB 4nM DNA

3) 1nM MB 960pM DNA

4) 500pM MB 400pM DNA

5) 100pM MB 80pM DNA

6) 50pM MB 40pM MB

7) 25pM MB 20pM DNA

• Run the following oligio-D samples

1) 20pM

2) 40pM

3) 60pM

4) 80pM

5) 100pM

## Calculations

MOLECULAR BEACON:

7.61μM (x) = 20nM (1000uL)

x = 2.62uL 7.61uM MB in 997.38uL PBS

20nM (x) = (30uL)(10nM)

15uL in 15uL PBS

20nM (x) = 2nM(100uL)

10uL in 90uL

2nM (x) = 1nM (100uL)

x = 50uL in 50uL PBS

200pM (100uL) = x (1nM)

x = 20uL in 80uL PBS

100pM(100uL) = (200pM)(x)

x = 50uL in 50uL buffer

50pM (30uL) = 100pM (x)

x = 15uL in 15uL

DNA:

5.31uM (500uL) = x (53.1uM)

x = 50uL in 450uL PBS

5.31uM (x) = 16nM (1000uL)

x = 3uL in 997uL PBS

16nM (x) = (8nM) (100uL)

x = 50uL in 50uL PBS

1.92nM (100uL) = x (8nM)

x = 24uL in 76 uL PBS

0.8nM (100uL) = x (1.92nM)

x = 41.7uL in 58.3uL

160pM (100) = x (800pM)

x = 20uL in 80uL

80pM (100uL) = x (160pM)

x = 50uL in 50uL

40pM (30uL) = x (80pm)

x = 15uL in 15uL PBS