GenElute™ PCR Clean-Up Procedure

Video: PCR Clean-Up Procedure

Product No. NA1020

The GenElute™ PCR Clean-up Kit is designed for rapid purification of single-stranded or double-stranded PCR amplification products (100 bp to 10 kb) from other components in the reaction, such as excess primers, nucleotides, DNA polymerase, oil and salts. DNA purification is achieved in a few easy steps. Purified DNA can be used in enzymatic reactions, conventional or automated sequencing, cloning and microarray analysis.

• Starting Amount - 100 µL or 10 µg of DNA
• Yield - Recovers up to 95% of PCR fragments between 100 bp to 10 kb
• Speed - Purify up to 100 µL or 10 µg of PCR amplified DNA in 5 minutes
• Flexible - DNA concentrated in 50 µL of buffer
• Economic - 40% more purifications than market leader

Sufficient for 70 purifications.

PCR Clean-Up Steps

1. Insert the binding column into a provided collection tube.

2. Add 0.5 mL of the column preparation solution to each miniprep column.

3. Centrifuge at 12,000 x g for 1 minute.

4. After 1 minute, discard the eluate.

5. Transfer PCR reaction mix into each tube.

6. Add 5 volumes of binding solution to 1 volume of the PCR reaction and mix.

7. Transfer the solution into the binding column.

8. Centrifuge at maximum speed for 1 minute.

9. After 1 minute, discard the eluate, but retain the collection tube.

10. Add 0.5 mL of diluted wash solution to the column.

11. Centrifuge at maximum speed for 1 minute.

12. After 1 minute, discard the eluate, replace the column back into the collection tube.

13. Centrifuge at maximum speed for 2 minutes (without adding additional wash solution.

14. After 2 minutes, discard any residual eluate and the collection tube. Transfer the column to a fresh 2 mL collection tube.

15. Add 50 µL of elution solution to the center of each column.

16. Centrifuge at maximum speed for 1 minute.

17. After 1 minute, discard the column.

18. PCR amplification products are now presented in the eluate.