FastDigest® HindIII

Kód produktu: FD0505 Kód výrobce: FD0505 Kód dodavatele: {1590198C-DFFF-4497-86BF-61D31C0A6B47} Výrobce: Life Technologies Czech Republic s.r.o.

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`5'...A↓A G C T T...3' 3'...T T C G A↑A...5'`

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Detailní popis

Thermo Scientific FastDigest enzymes are an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers.

The universal buffer allows rapid single-, double- or multiple DNA digestion within 5-15 minutes eliminating any need for buffer change or subsequent DNA clean-up steps. DNA modifying enzymes, such as Klenow Fragment, T4 DNA Ligase, alkaline phosphatases and T4 DNA Polymerase all have 100% activity in FastDigest Buffer. Therefore, enzymes for downstream applications can be directly added to the FastDigest reaction mix.

For additional convenience FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading of digestion reaction products on gels.

Short incubation times and optimal composition of the universal FastDigest Buffer eliminate star activity effects.

Features

100% activity of all FastDigest enzymes in the universal buffer
100% buffer compatibility with downstream applications
Complete digestion in 5-15 minutes
Direct loading on gels
No star activity
176 FastDigest enzymes available

Applications

Molecular cloning
Restriction site mapping
Genotyping
Southern Blot
Restriction fragment length polymorphism (RFLP)
SNP analysis

Note

The FastDigest Green Buffer offers the same high performance in DNA digestion and downstream applications as the colorless FastDigest Buffer. For applications that require product analysis by fluorescence excitation (e.g. concentration measurements in UV light) the colorless FastDigest Buffer is recommended.

For methylation sensitivity refer to product specifications.

Isoschizomers Search for commercial isoschizomers using REsearch.

Formulation	1 µL of enzyme (1 FDU) cleaves 1 μg of lambda DNA in 5 minutes at 37°C in 1X FastDigest Buffer.
Hazardous	No

Recommended Reaction Conditions	1X FastDigest buffer or 1X FastDigest Green buffer at 37°C. 1 µL of FastDigest HindIII is formulated to digest up to: 1 µg of lambda DNA in 5 minutes 1 µg of plasmid DNA in 15 minutes 0.2 µg of PCR product in 20 minutes 1 µg of genomic DNA in 10 minutes or 5 µg of genomic DNA in 30 minutes
Storage Condition	-20 C

Reaction conditions

Reaction temperature	Digestion time with 1 µL of FastDigest enzyme, min				bp from end of DNA required for complete digestion	Thermal inactivation	Incubation time without star activity, hours
Reaction temperature	Lambda, 1 µg/20 µL	Plasmid DNA, 1 µg/20 µL	PCR product, ~0.2 µg/30 µL	Genomic DNA, 1 µg/10 µL	bp from end of DNA required for complete digestion	Thermal inactivation	Incubation time without star activity, hours
37°C	5	15	20	10	3	80°C, 10 min	16

Methylation Effects

Methylation type	Sequence	Cleavage effect
EcoBI`(TGA(N)₈TGCT)`	`5'...TGm6AAGCTT(N)₃ TGCT...3'` `3'...AC TTCGAA(N)₃m6ACGA...5'`	Impaired

Number of recognition sites in DNA molecules

Lambda	ΦX174	M13mp18/19	pBR322	puc18/19	pUC57
6	0	1	1	1	1
pTZ19R/U	pTZ57R	pBluescriptIIKS(±)	pBluescriptIISK(±)	pACYC177	pACYC184
1	1	1	1	1	1

New sites generated by ligation

Newly generated recognition sites resulting from fill-in of 5'-overhang and self-ligation

Recognition Sequence	Newly generated sequence after reaction	Enzymes that cleave the newly generated sequence
`A^AGCTT`	`AAGCTAGCTT`	[AluI/FastDigest AluI] BspOI (BmtI)/FastDigest BmtI (BspOI) Cac8I [CviJI] FspBI (BfaI)/FastDigest BfaI (FspBI) NheI/FastDigest NheI [SetI]

Obsah balení

2500 rxns

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5'...A↓A G C T T...3' 3'...T T C G A↑A...5'

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