Calculate DNA Volume Needed for PCR Reactions
Target: 10 ng DNA per reaction
Good to go!
Add - μL of DNA + - μL of water to each PCR tube
Warning: Very Low Concentration!
Consider concentrating your DNA sample using vacuum centrifugation or ethanol precipitation.
Instructions:
1. Add calculated DNA volume to each PCR tube
2. Add calculated water volume to reach 10 μL total
3. Then add 15 μL of PCR master mix (from Master Mix Calculator)
4. Final reaction volume: 25 μL
| Concentration | DNA Volume | Water Volume | Total |
|---|---|---|---|
| 0.5 ng/μL | 20.0 μL | 0.0 μL | 20.0 μL |
| 1.0 ng/μL | 10.0 μL | 0.0 μL | 10.0 μL |
| 2.0 ng/μL | 5.0 μL | 5.0 μL | 10.0 μL |
| 5.0 ng/μL | 2.0 μL | 8.0 μL | 10.0 μL |
| 10.0 ng/μL | 1.0 μL | 9.0 μL | 10.0 μL |
| 20.0 ng/μL | 0.5 μL | 9.5 μL | 10.0 μL |
| 50.0 ng/μL | 0.2 μL | 9.8 μL | 10.0 μL |
Visual guide for typical DNA concentrations
Need >10 μL DNA
Concentrate sample
5-10 μL DNA needed
No water needed
1-5 μL DNA
Easy to pipette
<1 μL DNA
Precise pipetting