T4 DNA Ligase 200μL - Uniscience
Overview
T4 DNA Ligase catalyzes the formation of phosphodiester bonds between juxtaposed 5' phosphate and 3' hydroxyl termini in double-stranded DNA using ATP as a coenzyme. Both blunt and cohesive end DNA ligation, as well as single-stranded nick repair of DNA, RNA and DNA/RNA, are possible via the T4 DNA ligase. This enzyme is supplied in a buffer of 10 mM Tris-HCl (pH 7.5), 50 mM KCl, 10 mM 2-mercaptoethanol, 0.1 mM EDTA
and 50% glycerol.
Biological Source
- E. coli strain expressing a recombinant clone.
Molecular Weight
- 68kDa.
Requirements
Mg , ATP and DTT. The optimum concentration of Mg is 10mM. Mn may be substituted for Mg but is only 25% as effective as Mg .
Inhibition
50% inhibition by greater than 150mM NaCl (activity measured at nicks). Other inhibitors include 0.2M K , Cs , Li , NH and 1mM spermine.
Inactivation
- Heat to 70°C for 10 minutes.
Contents
- T4 DNA Ligase (200 U/μl).
- 10 X Ligation Buffer.
Applications
- Cloning of restriction fragments.
- Joining linkers and adapters to blunt-ended DNA.
Unit Definition
One unit of T4 DNA Ligase is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of λ DNA (5´ DNA termini concentration of 0.12 μM, 300- μg/ml) in a total reaction volume of 20 μl in 30 minutes at 16°C in 1x T4 DNA Ligase Reaction Buffer.
Storage
- Store at -20°C.
Storage Buffer
50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM DTT, 200 μg/ml BSA and 50% glycerol.
10X T4 Ligase Buffer
660mM Tris-HCl (PH7.6), 66mM MgCl , 100mM DTT, 1mM ATP.
Physical Purity
The purity is ≥90% as judged by SDS-polyacrylamide gel electrophoresis with Coomassie blue staining.
Standard Applications
We recommend using a 1:1, 1:3 or 3:1 molar ratio of vector:insert DNA when cloning a fragment into a plasmid vector. These ratios will vary with other types of vectors, for example, cDNA and genomic cloning vectors. The following example illustrates the conversion of molar ratios to mass ratios for a 3.0kb plasmid and a 0.5kb insert DNA
fragment.
Protocol
The following ligation reaction of a 3kb vector and a 0.5kb insert DNA uses a 1:1 vector:insert ratio. Typical ligation reactions use 100–200ng of vector DNA.
Warranty Information
LIMITED WARRANTIES
Limited Warranties are non-transferable. The following Limited Warranties are given to the original retail purchaser of the following Ashley Furniture Industries, Inc.Products:
Frames Used In Upholstered and Leather Products
Limited Lifetime Warranty
A Limited Lifetime Warranty applies to all frames used in sofas, couches, love seats, upholstered chairs, ottomans, sectionals, and sleepers. Ashley Furniture Industries,Inc. warrants these components to you, the original retail purchaser, to be free from material manufacturing defects.
Purchasing & Delivery
Before you make your purchase, it’s helpful to know the measurements of the area you plan to place the furniture. You should also measure any doorways and hallways through which the furniture will pass to get to its final destination.Picking up at the store
Shopify Shop requires that all products are properly inspected BEFORE you take it home to insure there are no surprises. Our team is happy to open all packages and will assist in the inspection process. We will then reseal packages for safe transport. We encourage all customers to bring furniture pads or blankets to protect the items during transport as well as rope or tie downs. Shopify Shop will not be responsible for damage that occurs after leaving the store or during transit. It is the purchaser’s responsibility to make sure the correct items are picked up and in good condition.Delivery
Customers are able to pick the next available delivery day that best fits their schedule. However, to route stops as efficiently as possible, Shopify Shop will provide the time frame. Customers will not be able to choose a time. You will be notified in advance of your scheduled time frame. Please make sure that a responsible adult (18 years or older) will be home at that time.In preparation for your delivery, please remove existing furniture, pictures, mirrors, accessories, etc. to prevent damages. Also insure that the area where you would like your furniture placed is clear of any old furniture and any other items that may obstruct the passageway of the delivery team. Shopify Shop will deliver, assemble, and set-up your new furniture purchase and remove all packing materials from your home. Our delivery crews are not permitted to move your existing furniture or other household items. Delivery personnel will attempt to deliver the purchased items in a safe and controlled manner but will not attempt to place furniture if they feel it will result in damage to the product or your home. Delivery personnel are unable to remove doors, hoist furniture or carry furniture up more than 3 flights of stairs. An elevator must be available for deliveries to the 4th floor and above.
Purchasing & Delivery
Before you make your purchase, it’s helpful to know the measurements of the area you plan to place the furniture. You should also measure any doorways and hallways through which the furniture will pass to get to its final destination.Picking up at the store
Shopify Shop requires that all products are properly inspected BEFORE you take it home to insure there are no surprises. Our team is happy to open all packages and will assist in the inspection process. We will then reseal packages for safe transport. We encourage all customers to bring furniture pads or blankets to protect the items during transport as well as rope or tie downs. Shopify Shop will not be responsible for damage that occurs after leaving the store or during transit. It is the purchaser’s responsibility to make sure the correct items are picked up and in good condition.Delivery
Customers are able to pick the next available delivery day that best fits their schedule. However, to route stops as efficiently as possible, Shopify Shop will provide the time frame. Customers will not be able to choose a time. You will be notified in advance of your scheduled time frame. Please make sure that a responsible adult (18 years or older) will be home at that time.In preparation for your delivery, please remove existing furniture, pictures, mirrors, accessories, etc. to prevent damages. Also insure that the area where you would like your furniture placed is clear of any old furniture and any other items that may obstruct the passageway of the delivery team. Shopify Shop will deliver, assemble, and set-up your new furniture purchase and remove all packing materials from your home. Our delivery crews are not permitted to move your existing furniture or other household items. Delivery personnel will attempt to deliver the purchased items in a safe and controlled manner but will not attempt to place furniture if they feel it will result in damage to the product or your home. Delivery personnel are unable to remove doors, hoist furniture or carry furniture up more than 3 flights of stairs. An elevator must be available for deliveries to the 4th floor and above.
