AMPure Cleaned PCR Product
07April10 Ethanol Cleaned Product (~30ul)
- Consumed 10ul of remaining Product
- Used Standard Agencourt AMPureXP Protocol
http://www.beckmangenomics.com/products/dna_purification_and_cleanup/agencourt_ampure_xp.html
- Final Elution step, only could retrieve ~30ul cleanly, minimal bead pick up.
Sample ID
|
Nucleic Acid Conc.
|
Unit
|
A260
|
A280
|
260/280
|
260/230
|
Sample Type
|
lco/hco 1 |
0.5 |
ng/µl |
0.01 |
0.005 |
1.91 |
3.78 |
DNA
|
lco/hco 9 |
1 |
ng/µl |
0.02 |
0.016 |
1.26 |
2.17 |
DNA
|
lco/hco 17 |
-0.1 |
ng/µl |
-0.003 |
-0.017 |
0.17 |
0.15 |
DNA
|
lco/hco 25 |
1.5 |
ng/µl |
0.03 |
0.008 |
3.98 |
0.84 |
DNA
|
lco/hco 33 |
1.2 |
ng/µl |
0.025 |
0.02 |
1.25 |
1.73 |
DNA
|
lco/hco 41 |
0.7 |
ng/µl |
0.014 |
-0.014 |
-1 |
1.66 |
DNA
|
lco/hco 49 |
1.7 |
ng/µl |
0.035 |
0.022 |
1.56 |
1.21 |
DNA
|
lco/hco 57 |
1.2 |
ng/µl |
0.024 |
0.013 |
1.82 |
1.07 |
DNA
|
lco/hco 65 |
1.5 |
ng/µl |
0.03 |
0.028 |
1.08 |
1.25 |
DNA
|
lco/hco 73 |
1.7 |
ng/µl |
0.034 |
0.014 |
2.5 |
1.19 |
DNA
|
'
|
10ul Ethanol Cleaned ->
|
Total PCR Product Recovery
|
|
|
AMPure Cleaned = 30ul
|
lco/hco 1 |
97 |
15
|
lco/hco 9 |
340 |
30
|
lco/hco 17 |
223 |
10
|
lco/hco 25 |
299 |
45
|
lco/hco 33 |
287 |
36
|
lco/hco 41 |
564 |
21
|
lco/hco 49 |
597 |
51
|
lco/hco 57 |
370 |
36
|
lco/hco 65 |
283 |
45
|
lco/hco 73 |
431 |
51
|
PEG Cleaned PCR Product
Clean remaining 07April10 Ethanol Cleaned PCR Product ~18ul with PEG.
To make PEG:
3g PEG 8000
2.19g NaCl
Bring up to 15ml with good water.
Lasts 30 days
Protocol:
1. Add 1:1 volume of PEG to PCR product.
2. Vortex briefly.
3. Warm at 37degrees for 15minutes.
4. Microcentrifuge for 15minutes at 4000rpm (cold is best).
5. Pipette off supernatant.
6. Wash pellet with 100ul 80% cold ethanol.
7. Spin for 10minutes at 4000rpm. Cold is best.
8. Pipette off ethanol.
9. Repeat steps 6-8 once or twice.
10. Dry down.
11. Resuspend in 10ul good water.
12. NanoDrop
1
|
0.2
|
ng/µl
|
0.004
|
-0.011
|
-0.38
|
0.17
|
DNA
|
50
|
9 |
0.4 |
ng/µl |
0.007 |
-0.013 |
-0.53 |
0.71 |
DNA |
50
|
17 |
0.5 |
ng/µl |
0.01 |
-0.016 |
-0.6 |
0.3 |
DNA |
50
|
25 |
2.8 |
ng/µl |
0.057 |
0.023 |
2.5 |
0.37 |
DNA |
50
|
33 |
0.4 |
ng/µl |
0.008 |
-0.011 |
-0.71 |
0.89 |
DNA |
50
|
41 |
1.5 |
ng/µl |
0.03 |
0 |
-1118.82 |
0.44 |
DNA |
50
|
49 |
4.1 |
ng/µl |
0.082 |
0.035 |
2.32 |
0.96 |
DNA |
50
|
57 |
2.7 |
ng/µl |
0.054 |
0.027 |
2 |
0.98 |
DNA |
50
|
65 |
0.9 |
ng/µl |
0.018 |
0.001 |
25.44 |
0.54 |
DNA |
50
|
73 |
1.9 |
ng/µl |
0.039 |
0.005 |
7.24 |
0.87 |
DNA |
50
|
76 |
1.8 |
ng/µl |
0.035 |
-0.003 |
-10.14 |
0.61 |
DNA |
50
|
|