Details
Two in one – DNA cleanup from agarose gels and PCR
The FastGene® Gel/PCR Extraction Kit is designed for the extraction of DNA from agarose gels and for the purification of PCR products. DNA fragments purified with FastGene® Gel/PCR Extraction Kits are ready for direct use in all common downstream applications, like sequencing, ligation and transformation, restriction digestion, microarray analysis, PCR and in vitro transcription.
Easy workflow
The FastGene® Gel/PCR Extraction Kit provides spin columns, buffers, and collection tubes for silica-membrane-based purification of DNA fragments from agarose gels and PCR products. With a simple and fast bind-wash-elute procedure you can purify DNA ranging from 15 bp to 10 kb with an elution volume of 20-50 µL.
Extra benefits
Each FastGene® Gel/PCR Extraction Kit includes 5 FastGene® Agarose Gel Band Cutters and 50 µL Midori Green Advance DNA stain (enough to stain 1 L of agarose gels). Everything you need to cut out your DNA fragments.
Applications
High recovery
The figure demonstrates that the FastGene® Gel/PCR Extraction Kit shows up to 90% of DNA recovery.
Figure: PCR fragments of 300 bp were purified from 40 μL of a PCR stock solution using FastGene® Gel/ PCR Extraction Kit and a competitor kit, according to manufacturers protocol. 5 μL of eluted DNA were analyzed on a 1.5% TAE agarose gel.
Extraction of small fragments
The DNA recovery from agarose gels using FastGene® Gel/PCR Extraction Kits is very high even for small fragments.
Figure: 10 µL of a PCR stock solution with a 100 bp amplicon were applied on a TBE agarose gel. After electrophoresis the DNA was purified using a competitor kit and the FastGene® Gel/PCR Extraction Kit, according to manufacturers protocol. DNA was eluted in 20 µL elution buffer and 10 µL of eluate was analyzed on a 2% TAE agarose gel.
Lane 1: Unpurified PCR fragment; M = Marker
Lane 2,3 and 4: Supplier M
Lane 5,6 and 7: FastGene® Gel/PCR Extraction Kit
Extraction of large DNA fragments
Background:
It is a well-known problem that the recovery of DNA fragments larger than 1 kb proves to be difficult and leads to the loss of large amounts of DNA. In this AppNote the FastGene® Gel/PCR Extraction Kit was used for the isolation of two DNA bands after restriction digestion.
Results/Conclusion:
Both fragments show a good recovery rate after extraction. The customer also highlighted the fast preparation, easy handling, high recovery rate for large fragments and the unproblematic performance in downstream applications.
Method:
A 6.9 kb large plasmid was digested with a restriction enzyme. The restriction digest was analysed by agarose gel electrophoresis at 100 V for 20 min. The 0.7% agarose gel was produced using 1x TAE buffer (Figure 1). The target fragments were excised out of the gel and transferred in a 1.5 mL tube. The fragments were purified with the FastGene® Gel/PCR Extraction Kit. 100 ng of each purified DNA fragment were electrophoresed again at 100 V for 20 min (Figure 2).
Results/Conclusion:
Both fragments show a good recovery rate after extraction. The customer also highlighted the fast preparation, easy handling, high recovery rate for large fragments and the unproblematic performance in downstream applications.
FAQ
Is the GP1 buffer of this kit available separately like for the Mini Kit Buffer Set?
A: We do not sell GP1 separately or as a Buffer Set.
What is the composition of the GP3 Buffer (does it contain EDTA or EtOH)?
The elution buffer GP3 contains just 10 mM Tris Buffer pH8 and no EtOH or EDTA.
Can I use water for the elution and what pH has the water to have?
You can use water but we would recommend to use the GP3 buffer. The pH should be between 7 and 8. If you buy water this should be OK. Please check the elution efficiency. If this is not OK check the pH of the water or use GP3.
Only logged in customers may leave a review.
Dr. Ivana Primorac, Universität Duisburg-Essen, Department of Molecular Genetics –
For a considerably lower price, this kit allows purification of PCR products, as well as recovery of DNA from an excised gel, with yields comparable to the more expensive kits on the market. The workflow is the same/very similar to those of other PCR/Gel extraction kits. I can only recommend it.
Verified User