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11635379001 Roche

Rapid DNA Ligation Kit

Synonym: dna ligation kit, rapid, ligation kit, rapid dna

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Properties

Quality Level   100
usage   sufficient for 40 reactions (DNA ligation)
mfr. no.   Roche
shipped in   dry ice
storage temp.   −20°C

Description

General description

The Rapid DNA Ligation Kit contains all reagents necessary for ligation. No additional reagents or additives are required. The kit can rapidly ligate DNA with either blunt or sticky ends at +15 to +25°C. Depending on the DNA concentration in the reaction, the ligation products will be either circular (if the DNA concentration is low) or concatemeric (if the DNA concentration is high). Ligated DNA is suitable for direct use in transformation experiments.

Note: Electroporation can be performed after ligation in combination with the High Pure® PCR Product Purification Kit.

Specificity

DNA with either blunt or sticky ends (≤200 ng).
Heat inactivation: 10 min at 65 °C (T4 Ligase)
Inactivation of T4 Ligase is necessary only if the ligation reaction mixture is to be used in experiments other than transformation assays.
Note: Heat inactivation of the ligation reaction mixture before transformation causes the number of transformed colonies to decrease drastically (> factor of 20).
Star activity: Star activity should also be avoided. When double digests are performed with enzymes prone to star activity, consult the Roche Restriction Enzyme Poster (or the Roche Applied Science Catalog) to select the optimal buffer for a given enzyme combination. Whenever possible use Buffer H for double digests.

Application

The Rapid DNA Ligation Kit covalently joins either sticky-ended or blunt-ended DNA fragments, for example, for:
• Cloning into plasmid or phage vectors.
• Adding linkers to plasmids or other DNAs.
• Recircularizing linear DNA.

Features and Benefits

• Fast: ligation takes only 5 minutes.
• Convenient: ligation is performed at +15°C to +25°C.
• Easy: kit contains all buffers and enzymes needed.
• Flexible: kit can be used for all common ligation reactions.
• Reliable: simple procedure and ready-to-use reagents ensure accuracy and reproducibility.

Packaging

1 kit containing 3 components.

Preparation Note

Incubation time: 5 minutes
Molar ratio of DNA to be ligated: The molar ratio of vector DNA to insert DNA in the standard ligation reaction is normally 1:3. However, when appropriate, you may use other ratios. For example:
If the two DNAs differ greatly in length, try a 1:1 or 1:2 ratio
If the ends of the two DNAs are blunt, try a 1:5 ratio

Other Notes

The new Rapid DNA Dephos & Ligation Kit contains all the reagents of the Rapid DNA Ligation Kit plus our new rAPid Alkaline Phosphatase.

Legal Information

High Pure is a registered trademark of Roche

Kit component only

Description

Product #

Add to Cart

Dilution Buffer 5x concentrated    
T4 DNA Ligation Buffer 2x concentrated    
T4 DNA Ligase 5 U/μl    
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
Flash Point(F) 
does not flash
Flash Point(C) 
does not flash

Documents

Certificate of Analysis (COA)

Please Enter a Lot Number
Protocols & Articles

Protocols

Rapid DNA Ligation Kit Protocol

To perform a successful cloning experiment, use these handling steps as guidelines for using the Rapid DNA Ligation Kit.
Keywords: Antibiotics, Buffers, Cloning, Genetic, Peptide synthesis, Polymerase chain reaction, Precipitation, Purification, transformation

Peer-Reviewed Papers
15

References

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