T4 DNA Ligase

Intact Genomics T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5'-phosphate and 3'-hydroxyl termini in duplex DNA or RNA.  This enzyme joins DNA fragments with either cohesive or blunt termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.
 

Quality Control 
Quality control is performed following the production of each new lot of product to ensure that it meets the quality standards and specifications designated for the product. Each lot is repeatedly compared side-by-side with leading competitors to ensure our products outperform the competitor before product launching. 

Intact Genomics T4 DNA Ligase displays up to 3-5X higher ligation efficiency than the nearest competitor.

Product Source
E. coli strain expressing a recombinant clone

Quality Control 
Quality control is performed following the production of each new lot of product to ensure that it meets the quality standards and specifications designated for the product. Each lot is repeatedly compared side-by-side with leading competitors to ensure our products outperform the competitor before product launching. 

Contents & Storage

1. T4 DNA Ligase
2. 10x T4 DNA Ligase Reaction Buffer (w/o ATP)
3. 10 mM ATP

Store all contents at -20 °C.

Storage Buffer 
50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25 °C

10x T4 DNA Ligase Reaction Buffer (w/o ATP) 
500 mM Tris-HCl, 100 mM MgCl, 100  mM DTT, pH 7.5 @ 25 °C

Note
10x T4 DNA ligase buffer does not contain ATP. You need to add ATP separately.

Unit Definition 
One Weiss unit is defined as the amount of enzyme required to convert 1 nmol of 32P from pyrophosphate into Norit-absorbance material in 20 minutes under standard assay conditions.

Protocol

1. Set up reaction buffer in a microcentrifuge tube on ice. Use a molar ratio of 1:3 vector to insert DNA.

1. Gently mix the reaction and centrifuge briefly.
2. For cohesive ends, incubate 16 °C for overnight or at room temperature for 30 min.
3. For blunt ends, incubate 16 °C for overnight or at room temperature for 2 hrs.
4. Heat inactivate at 70 °C for 15 min.
5. Cool on ice and transform 2 µl of the reaction into 50 µl competent cells.

ig-Fusion Cloning Kit 
Intact Genomics propriety ig-Fusion cloning technology is a simple, rapid and highly efficient cloning kit which allows to directly clone any PCR product(s) to any linearized expression vector at any site. The PCR fragments can be generated by Intact Genomics high fidelity Pfu DNA polymerase or other high-fidelity DNA polymerases, with primers having 15 to 18 bases of homology at their linear ends to where the product need to fuse. The linearized vector can be generated by PCR or restriction enzymes. The kit is so robust that multiple DNA fragments can be assembled simultaneously and cloned into one construct in a single reaction step within short times (usually 10-30 min) with more than 95% cloning efficiency.

Benefits

• Clone any insert at any site within any vector
• Restriction enzyme and phosphatase free system
• Joining multiple large fragments at once
• Precise insertion at a desired orientation
• Rapid and high efficiency with > 95% positive clones

Quality Control 
Quality control is performed following the production of each new lot of product to ensure that it meets the quality standards and specifications designated for the product. Each lot is repeatedly compared side-by-side with leading competitors to ensure our products outperform the competitor before product launching. 

Contents & Storage

• 5x ig-Fusion enzyme premix: -20 °C
• 2x PCR premix: -20 °C
• High efficiency competent cells: -80 °C
• Recovery medium:4 °C or -20 °C

Protocol
1. Linearize the vector by restriction enzyme digestion or inverse PCR and purify the product with spin column.
2. Design PCR primers for the gene of interest with 15 to 20 bp at 5'-extensions that are complementary to the ends of the linearized vector.
3. Amplify the gene of interest with Intact Genomics 2x PCR premix or any other high-fidelity DNA polymerase. Run the PCR product on an agarose gel to determine the integrity of the PCR product.
4. Purify the PCR product with spin column.
5. Set up the ig-Fusion cloning reaction as follows: Insert and vector molar ratio 3:1 produce the highest number of colonies.  

6.  Mix the reaction mixture thoroughly.
7.  Incubate the reaction mixture at 50 °C for 10-30 min, then place on ice. Number of colonies depend on the incubation time, insert size and number of inserts need to clone.
8.  Use 2.0 µl of the reaction mixture and transform into high efficiency ig 10B chemical or electroporation competent cells (included). To get the maximum number of colonies, we recommend to use ig 10B electrocompetent cells (Cat # 1212).

ENDURO™ Semi Dry Blotter

• Western, Southern, and Northern Blots
• Rapid transfer times
• Uniform head dispersion
• Variable gel thickness
• Minimal buffer volume

The ENDURO™ Semi-Dry Blotter provides great flexibility in a lab since it can be used for all types of blotting: Western, Southern, and Northern. Blotting with the ENDURO™ Semi Dry Blotter typically can be done in 15 to 30 minutes making it a fast and easy way to do all your blotting applications. Set up procedures are easy and economical with little buffer needed. The screw down lid adjusts to varying gel thicknesses and sizes while the platinum coated electrodes provide uniform pressure ensuring even transfers. To provide your lab with total blotting solutions use with our ENDURO™ 250V, 3000mA high current power supply.

Specifications

Semi-Dry Blotting Systems, 10cm x 10cm

• Small footprint for benchtop use
• Preset and manual controls for ultraviolet or time exposures. Preset exposure delivers 120,000 microjoules or five minutes of exposure
• Microprocessor measures and control UV output, ensuring maximum energy efficiency
• Large LED display and touch panel which displays time or energy settings
• Window on the door allows viewing of the process yet blocks out the UV radiation
• Select from models with 302nm midrange UV, 365nm longwave UV or 254nm shortwave UV Interior: 12 deep x 10 wide x 5 high inches (305 x 254 x 127mm)
• Overhead UV supplies uniform UV
• Shortwave UV model is available with choice of door (CL-1000) or drawer (CX-2000) styles
• Built-in UV sensor is calibrated at the UVP factory to ensure accuracy and reliability
• Crosslinking for attaching nucleic acids to a membrane takes seconds as compared to oven baking

CL-1000 Dimensions: Exterior: 13.7 deep x 15.6 wide x 8.75 high inches (248 x 396 x 222mm)

CX-2000 Dimensions:
Exterior: 17.5"W x 15"D x 9.75"H (445 x 381 x 248mm)
Interior: 11.4"W x 11.3"D x 3.6"H (290 x 287 x 91mm)
Depth of sample tray 11.25 Width of sample tray 11.5
Sample surface to lamp approximately 3.5 in. (89mm)

1 year warranty

• Lamps are lightweight.
• Models available include shortwave, longwave UV or a combination shortwave/longwave version
• Combination version comes with a snap-on wavelength selector for switching between wavelengths
• Use these lamps with the J-129 Lamp Stand (pictured) or the C-10 Cabinet
• 1 year warranty
  
  Handheld Lamp Dimensions: 
  14.9"L x 3.2"W x 2.5"H
  Lamp Stand Dimensions: 9.5"H x 13"L

Shortwave XX Series Lamps use ultraviolet radiation as a reliable and effective tool for sterilization and sanitation applications in hospitals, laboratories, pharmaceutical companies, and clinics. Work surfaces and equipment can become contaminated with bacteria which may be passed through the air or by hand infecting research projects and workers. In addition to bacteria, yeast, mold spores, germs, and viruses may also be present. Shortwave XX Series UV Lamps can kill up to 99% of these organisms. Decontaminate work surfaces with UV prior to PCR to reduce air-borne contamination during DNA sequencing. UV sterilization is particularly useful for temperature sensitive applications due to low operating lamp temperatures. 

Shortwave Lamps can be used in an upward facing position for purifying the air or in a downward position for sanitizing work surfaces. Hanging the lamp (brackets provided) reduces the space required on laboratory benchtops. Longwave UV Bulbs (365nm) also available for all models. 1 year warranty

Dimensions
XX-15: 19.75"L x 6"W x 4.25"H
XX-20: 24"L x 6"W x 4.25"H
XX-40: 49.6"L x 6"W x 4.25"H
Exposure Stand: 20"L x 6"W x 13"H

Large Tank Blotter with two cassettes, 20 x 20cm

Semi-Dry Blotting Systems, 20cm x 20cm