Make a Standard DNA Plate
This step creates a Standard DNA plate with serial dilutions of stock Lambda DNA in the wells of column 1.
| 1. | Add stock Lambda DNA to well A1 in the Standard DNA plate and dilute to 75 ng/µl in a final volume of 233.3 µl. Pipette up and down several times to mix. |
| – | Use the following formula to calculate the amount of stock Lambda DNA: |
| – | Dilute the stock DNA in well A1 using the following formula: |
| 2. | Add 66.7 µl 1X TE to well B1. |
| 3. | Add 100 µl 1X TE to the remaining wells of column 1. |
Dilution of Stock Lambda DNA Standard
| 4. | Transfer 133.3 µl Lambda DNA from well A1 to well B1. Pipette up and down several times to mix. |
| 5. | Transfer 100 µl from well B1 to well C1. Pipette up and down several times to mix. |
| 6. | Repeat the sequential transfer of 100 µl for wells D1, E1, F1, and G1. Do not transfer from well G1 to H1. Well H1 serves as the blank 0 ng/µl Lambda DNA. |
|
Row-Column |
Concentration (ng/µl) |
Final Volume in Well (µl) |
|---|---|---|
|
A1 |
75 |
100 |
|
B1 |
50 |
100 |
|
C1 |
25 |
100 |
|
D1 |
12.5 |
100 |
|
E1 |
6.25 |
100 |
|
F1 |
3.125 |
100 |
|
G1 |
1.5262 |
200 |
|
H1 |
0 |
100 |
Serial Dilutions of Lambda DNA
| 7. | Cover the Standard DNA plate with a cap mat. |
