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RNA Ligases

NEB offers a selection of products for the ligation of single-stranded or double-stranded RNA to either RNA or DNA. Our RNA Ligase Selection Chart lists all of the ligases available for RNA, along with their recommended application.  

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Protocols for RNA Ligases
Reagents for RNA Sample Preparation
NEB offers a wide variety of reagents to facilitate front-end processing of RNA for downstream applications including next-generation sequencing, library construction and microarrays.
RNA Polymerase Selection Chart
NEB offers RNA polymerases that can be used for in vitro synthesis of RNA for a wide variety of downstream applications. Additional polymerases are offered for the generation of untemplated homoribopolymeric tails for reverse transcription or labeling.
RNA Ligase Selection Chart
NEB offers a variety of ligases for RNA research with unique specificities.
cDNA Synthesis Selection Chart
NEB offers several reagents for cDNA synthesis for use in applications including qPCR and qRT-PCR. For your convenience, reagents are available as kits or standalone products, depending on your needs.
Applications of T4 RNA Ligase 2, Truncated K227Q
  • Ligate a pre-adenylated DNA or RNA sequence tag to the 3´-end of any RNA
  • Join a single stranded adenylated primer to small RNAs for cDNA library creation
  • Join a single stranded adenylated primer to RNA for directional cDNA library construction
Advantages of the 5' DNA Adenylation Kit
  • Enzymatic 5´-adenylation of ss-DNA linkers for next gen sequencing
  • One step reaction gives quantitative adenylation.
  • Simpler than existing chemical and enzymatic methods.
  • Reduces need for purification of reaction product
  • 65°C reaction temperature reduces secondary structural concerns
  • Easily scalable from pmol to μmol range
Lower Undesired Ligation Products with T4 RNA Ligase 2 Truncated K227Q
1) T4 RNA Ligase 1; 2) T4 RNA Ligase 2, truncated; 3) T4 RNA Ligase 2, truncated K227Q. A 5´-phosphorylated 21-mer ssRNA was incubated with a 3´-blocked 17-mer preadenylated DNA adapter in a 1:2 molar ratio. Reactions contained RNA ligases as indicated. Ligation products were resolved by 15% denaturing PAGE and visualized with SybrGold nucleic acid stain.
Reported Activities and Applications for T4 Ligases
The ligation reactions depicted have been reported, but may require optimized reaction conditions.
Relative Efficiency of T4 Ligases on Nicked Subtrates*
+++ rate is greater than 10
++ rate is 1 - 10
+ rate is .1 - 1
– rate is less than 0.1

*Courtesy of Dr. Richard Bowater, University of East Anglia, Norwich, UK. Original graph appears in Bullard D. and Bowater R. (2006) Biochem J., 398, 135–144.
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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.


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