FastDigest® enzymes are an advanced line of restriction enzymes for rapid DNA digestion. All FastDigest® enzymes are 100% active in the universal FastDigest® and FastDigest® Green buffers and are able to digest DNA in 5-15 minutes. This enables any combination of restriction enzymes to work simultaneously in one reaction tube and eliminates the need for sequential digestions. As an added convenience, the FastDigest® Green Buffer allows for direct loading of the reaction mixture on a gel.
FastDigest® enzymes can be used to digest plasmid, genomic and viral DNA as well as PCR products and do not show star activity even in prolonged incubations.
The FastDigest® line is ideal for use in applications that require high purity reaction components, performance reliability and simple reaction set-up.
Universal FastDigest® Buffer
Figure 1. FastDigest® Green Buffer.
Reaction mixture containing FastDigest® Green buffer:
1 – before electrophoresis
2 – after electrophoresis
Two versions of the universal FastDigest® buffer are supplied with each enzyme: 10X FastDigest® Buffer and 10X FastDigest® Green Buffer. All FastDigest® enzymes are 100% active in both FastDigest® and FastDigest® Green buffers. Enzymes used in common downstream applications such as ligation, blunting and dephosphorylation also have 100% activity in both buffers. As an added convenience, the FastDigest® Green Buffer contains a density reagent and two tracking dyes that allow for direct loading of the reaction mixture on gels. The blue dye migrates with 3-5 kb DNA fragments in 1% agarose gel and has an excitation peak at 424 nm while the yellow dye migrates faster than 10 bp DNA fragments in 1% agarose gel and has an excitation peak at 615 nm.
The presence of the dyes in the FastDigest® Green buffer does not interfere with DNA digestion or with downstream applications but may hinder digestion product analysis by fluorescence excitation. For such applications we recommend use of the colorless FastDigest® Buffer.
- 100% activity of all FastDigest® enzymes in the new green universal digestion buffer.
- 100% buffer compatibility with all downstream applications.
- Complete digestion in 5-15 minutes.
- Direct loading of reaction mixture on gels.
- Fast clone analysis.
- Fast preparation of DNA for cloning.
- Digestion of PCR products.
- Fast RFLP genotyping.
- Digestion of difficult-to-cleave DNA.
Figure 2. Five minute triple digestion followed by ligation of plasmid DNA in the universal FastDigest® and FastDigest® Green buffers.
M – GeneRuler™ Express DNA Ladder (#SM1551)
C – undigested plasmid DNA control
1 – plasmid DNA triple digested with FastDigest® EcoRI, FastDigest® KpnI and FastDigest® SmaI in FastDigest® buffer
2 – the digestion and ligation reaction mixture in FastDigest® buffer
3 – plasmid triple digested with FastDigest® EcoRI, FastDigest® KpnI and FastDigest® SmaI in FastDigest® Green buffer
4 – the digestion and ligation reaction mixture in FastDigest® Green buffer