| Field | Specification |
|---|---|
| Mfr No | |
| Product Type | |
| Shipping | |
| Source | Recombinant (E. coli) |
| Storage |
PBCV DNA Ligase, also known as SplintR ligase, PBCV-1 DNA Ligase, or Chlorella virus DNA ligase, is an ATP-dependent DNA ligase that efficiently catalyzes the ligation reaction of two adjacent DNA strands that are complementary to a single stranded RNA. In these two adjacent DNA strands, the one providing the 3' hydroxyl group is called the acceptor DNA strand, while the one providing the 5' phosphate group is called the donor DNA strand. During this ligation process, a complementary RNA strand acts as a "splint" or "scaffold" for the two DNA strands. The activity of PBCV DNA Ligase is much higher than that of traditional T4 DNA ligase and shows a strong affinity for DNA substrates with RNA as a splint (apparent Km = 1 nM, where a smaller Km value indicates a stronger affinity). It can detect unique RNA species at the subnanomolar level in complex mixtures, making it suitable for target enrichment processes in applications such as high-sensitivity RNA detection, next-generation sequencing, and molecular diagnostics.
Features
No residual nuclease, endonuclease, RNase, or phosphatase
Applications
Detection of microRNAs and other small RNAs based on probe method
Connecting single-stranded DNA through "clamp" fixation by complementary RNA sequences
Using DNA probes to achieve the detection of specific RNA
Detection of SNP or Alternative Splicing
RASL-seq analysis
Specifications
|
Source |
The DNA Ligase gene from PBCV-1 originating from Chlorella virus expressed in E.coli |
|
Molecular Weight |
34 KD |
|
Concentration |
25 U/µL |
|
Unit Definition |
One unit is defined as the enzyme amount required to achieve at least 50% completion of the ligation of 2 pmol FAM-labeled DNA:RNA hybrid substrate within 15 min at 25℃ in 1×PBCV Ligase buffer |
|
Inactivation Conditions |
65℃, 20 min |
Glycerol Content |
Contains Glycerol |
Components
|
Component No. |
Name |
14962ES80 |
14962ES90 |
|
14962-A |
PBCV DNA ligase (25 U/µL) |
50 µL |
200 µL |
|
14962-B |
10 x PBCV Ligase buffer |
1 mL |
1 mL |
Shipping and Storage
The products should be stored at -25℃ ~ -15℃ for 2 years.
Store this enzyme at -20°C and avoid repeated freeze-thaw cycles to preserve catalytic activity. The product is shipped Dry Ice and remains stable for up to one year from the date of manufacture when stored under recommended conditions. Aliquoting the stock solution into single-use volumes is recommended for enzymes used infrequently to minimize thermal cycling of the bulk stock.
Detection of microRNAs and other small RNAs based on probe method Connecting single-stranded DNA through "clamp" fixation by complementary RNA sequences Using DNA probes to achieve the detection of specific RNA Detection of SNP or Alternative Splicing RASL-seq analysis. Always verify compatibility with your specific template, buffer, and downstream workflow.
One unit (U) is defined as the amount of enzyme that catalyzes the ligation of greater than 95% of HindIII-linearized lambda DNA fragments into high-molecular-weight concatemers in 30 min at 16°C in a 20 µL reaction volume.
This enzyme is produced as Recombinant (E. coli) and supplied as a Research Use Only (RUO) reagent. Each lot is subjected to activity assay, purity assessment by SDS-PAGE, and functional validation prior to release. A Certificate of Analysis (CoA) and Safety Data Sheet (SDS) are available on request.
Standard T4 DNA ligase requires overnight ligation at 16°C for optimal efficiency with cohesive ends. Quick Ligase formulations contain optimized buffer systems and enhanced enzyme concentrations that achieve >90% ligation efficiency within 5–15 minutes at room temperature, significantly accelerating cloning workflows without compromising junction fidelity.
Yeasen Biotechnology supports custom enzyme solutions across multiple service lines — from GMP-grade bulk supply to directed enzyme engineering. Contact BioHippo to discuss requirements and initiate a project inquiry.
▶ GMP-Grade & Bulk Supply
Select Yeasen enzymes are available in GMP grade, manufactured in an ISO 13485-certified UCF.ME™ ultra-clean molecular enzyme facility with FDA Drug Master File (DMF) support.
- GMP-grade release testing and CoA documentation
- ISO 13485-certified production facility
- Scalable from milligram to multi-gram quantities
- Consistent lot-to-lot activity specifications
▶ Glycerol-Free & Custom Formulation
Glycerol-free enzyme formats are available for applications requiring lyophilization compatibility, liquid handling automation, or direct IVD master mix integration.
- Glycerol-free liquid format (standard and custom buffers)
- Lyophilization-ready enzyme preparation
- Custom reaction buffer optimization for specific assay conditions
- Compatible with freeze-drying workflows for point-of-care formats
▶ Molecular IVD RDC Service
Yeasen's Research and Development Contracting (RDC) team delivers end-to-end solutions for molecular diagnostic product development, covering enzyme selection through clinical validation support.
- Enzyme selection and performance matching
- Primer/probe design and reaction buffer optimization
- Sensitivity, specificity, and precision validation studies
- Stability studies and SNP evaluation
- Instrument platform compatibility assessment
▶ ZymeEditor™ Enzyme Engineering
Yeasen's proprietary ZymeEditor™ directed evolution and rational design platform enables the development of custom enzyme variants with tailored performance characteristics not available in off-the-shelf products.
- Directed evolution for enhanced thermostability, processivity, or fidelity
- Rational design for altered substrate specificity or cofactor requirements
- Library screening from Yeasen's proprietary enzyme variant collection
- Scale-up to commercial quantities upon candidate confirmation
ⓘ Customization services are fulfilled by Yeasen Biotechnology. Lead times and minimum order quantities vary by service type. Contact BioHippo for project scoping and pricing.
