Basic Description

Storage Temp

Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

This product requires cold chain shipping. Ground and other economy services are not available.

Product Description

T4 DNA Ligase is isolated and purified from Escherichia coli expressing the T4 DNA Ligase gene after induction and expression. It catalyzes the binding reaction of adjacent DNA strands with 5 'phosphate groups and 3' hydroxyl groups via phosphodiester bonds. This enzyme can catalyze the connection of flat or sticky end DNA, repairing single stranded incisions in double stranded DNA, RNA, and DNA/RNA hybridization, but has no activity for single stranded nucleotides.

T665705	Component	100 U	500 U	5 KU	Storage
T665705A	T4 DNA Ligase, 5 U/μL	20 μL	100 μL	1 mL	-20℃. Avoid freeze/thaw cycle.
T665705B	10×Ligation Buffer	150 μL	750 μL	5×1.5 mL	-20℃. Avoid freeze/thaw cycle.
T665705C	50% PEG Solution	150 μL	750 μL	5×1.5 mL	-20℃. Avoid freeze/thaw cycle.

Preparation and important precautions before the experiment:
The final dosage of T4 DNA Ligase should not exceed the recommended dosage, otherwise it will affect the connection efficiency.
2. PEG can greatly improve the connection efficiency of the flat end. We recommend adding a final concentration of 5% PEG Solution to improve the connection efficiency
The connection efficiency of the high flat end.
3. To improve conversion efficiency, it is recommended that the amount of connecting products added should not exceed 10% of the volume of receptive cells
4. Due to the presence of glycerol in T4 DNA ligase, which is viscous and prone to wall sticking, it is recommended to briefly centrifuge the liquid before use
Collect to the bottom of the tube, and when sampling, try not to let the nozzle go too deep into the liquid level to avoid sticking to the nozzle and causing losses
Usage:
i Connection at the adhesive end
1. Reaction system:

Component	20 μL Reaction system	Final concentration
Linear vector DNA	X μL	20-100 ng
Inserting DNA fragments	Y μL	Insertion fragments: vector 1:1-5:1
10×Ligation Buffer	2 μL	/
T4 DNA Ligase 5 U/ μL	0.2 μL	1 U
ddH20	Add to 20 μL	20 μL

2. Vortex oscillation, instant centrifugation, collects the solution on the pipe wall to the bottom of the pipe.
3. Reaction conditions: Incubate at 22C for 10 minutes.
4. Instantly centrifuge and collect the solution on the tube wall to the bottom of the tube. Incubate at 65C for 10 minutes or 70C for 5 minutes to inactivateT4 DNA Ligase
5. Take 5pL of the connecting product and heat shock transform it into 5uL of competent cells, or take 15pL of the connecting product and shock transform it into 50pL of competent cells Cells.
Attention: If electric shock conversion is required, it is recommended to use ethanol precipitation method to remove T4 DMA Ligase before conducting electric shock conversion
Ii. Connection at the flat end
1. Reaction system:

Component	20 μL Reaction system	Final concentration
Linear vector DNA	X μL	20-100 ng
Inserting DNA fragments	Y μL	Insertion fragments: vector 1:1-5:1
10xLigation Buffer	2 μL	/
T4 DNA Ligase，5 U/uL	1 μL	5 U
50%PEG Solution	2 L	5%
ddH20	Add to 20 μL	20 μL

2. Vortex oscillation, instant centrifugation, collects the solution on the pipe wall to the bottom of the pipe.
3. Reaction conditions: Incubate at 22C for 1 hour.
4. Instantly centrifuge and collect the dissolved waves on the tube wall to the bottom of the tube. Incubate at 65C for 10 minutes or 70C for 5 minutes to inactivate T4 DNA ligase
5. Take 5L of connecting products and heat shock convert 50pL of competent cells, or take 1-2L of connecting products and shock convert 50pL of competent cells.
Note: If electroconversion is required, it is recommended to use ethanol precipitation method to remove T4 DNA ligase before electroconversion
iii Self cyclization of linear DNA
1. Reaction system:

Component	50 μL Reaction system	Final concentration
Linear vector DNA	X μL	5-50 ng
10xLigation Buffer	5 μL	/
T4 DNA Ligase，5 U/uL	1 μL	5 U
ddH20	Add to 50 μL	50 μL

2. Vortex oscillation, instant centrifugation, collects the solution on the pipe wall to the bottom of the pipe.
3. Reaction conditions: Incubate at sticky end 22C for 10 minutes, and incubate at flat end 22C for 1 hour.
4. Instantly centrifuge and collect the solution on the tube wall to the bottom of the tube. Incubate at 65C for 10 minutes or 70C for 5 minutes to inactivate T4 DNA ligase
5. Take 5pL of connecting products and heat shock them to convert 5pL of competent cells, or take 1-2pL of connecting products and shock them to convert 50pL of competent cells.
Note: If electroconversion is required, it is recommended to use ethanol precipitation method to remove T4 DNA ligase before electroconversion

Certificates（CoA,COO,BSE/TSE and Analysis Chart)

C of A & Other Certificates(BSE/TSE, COO):

Analytical Chart:

Documents & Articles

t665705_t4 dna ligase.pdf

PCR Laboratory Contamination Prevention and Efficient Reagent Application: —Optimizing Experimental Steps and Choosing

Categories: Technical articles

Tags:

Buffer PCR DNA polymerase PCR reagents and kits PCR and amplification Cloning and gene expression

Citations of This Product

How to Cite Aladdin Scientific Products?

1. Yiming Zhang, Zhi Chen, Songrui Wei, Jing Wang, Yujun Zhang, Huiling Lin, Hai Fu, Yingxia Liu, Qi Gao, Han Zhang, Zhongjian Xie. (2025) CRISPR CLAMP: Attomolar level of multiple miRNAs. CHEMICAL ENGINEERING JOURNAL, (161990).

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SKU	Size	Availability	Price
T665705-100U	100U	Available within 8-12 weeks(?) Production requires sourcing of materials. We appreciate your patience and understanding.	$49.90
T665705-500U	500U	Available within 8-12 weeks(?) Production requires sourcing of materials. We appreciate your patience and understanding.	$199.90
T665705-5KU	5KU	Available within 8-12 weeks(?) Production requires sourcing of materials. We appreciate your patience and understanding.	$1,499.90

T4 DNA Ligase