Properties of DNA and RNA Ligases
NEB offers many different kinds of DNA and RNA ligases for your needs. This chart will help you understand the differences between them.To select a ligase based upon the type of substrate present, please visit Substrate-based Ligase Selection Chart. In addition, the DNA Ligase Selection Chart helps you select the optimal DNA ligase for various applications.
| Ligase | Product Number | Recommended Reaction Temp. (Useful Range) |
Heat inact. | Co- factor |
Main Applications | Other notes |
|---|---|---|---|---|---|---|
| T4 DNA Ligase | M0202S/L | 25°C (4-37°C) |
Y (65°C) |
ATP | Ligation of nicks in dsDNA and joining dsDNA fragments with complementary overhangs > 2 bases in length. | T4 DNA Ligase can ligate other substrates with reduced efficiency, including: nicks containing mismatches and 1 or 2 nt gaps; dsDNA fragments with blunt ends and single-base overhangs; and the 3' end of RNA to a 5'-pDNA when annealed to a DNA complement. |
| T4 DNA Ligase, High Concentration | M0202T/M | 25°C (4-37°C) |
Y (65°C) |
ATP | Ligation of nicks in dsDNA and joining dsDNA fragments with complementary overhangs > 2 bases in length. Routine cloning. | 5X enzyme concentration over standard T4 DNA Ligase, good for increasing reaction rates or for bulk users. See additional notes for T4 DNA Ligase. |
| Hi-T4™ DNA Ligase | M2622S/L |
25°C (4-50°C) |
Y (65°C) |
ATP | Ligation of nicks in dsDNA and joining dsDNA fragments with complementary overhangs > 2 bases in length. | Hi-T4 DNA Ligase displays increased thermotolerance compared to T4 DNA Ligase. Hi-T4 DNA Ligase can withstand temperatures as high as 45°C for extended periods (up to 72 hours) without any significant loss in activity. |
| Salt-T4® DNA Ligase | M0467S/L |
25°C (4-37°C) |
Y (65°C) |
ATP | Ligation of nicks in dsDNA and joining dsDNA fragments with complementary overhangs > 2 bases in length. | Salt-T4 DNA Ligase will join cohesive end termini at salt concentrations as high as 300 mM without any loss in activity. Salt-T4 DNA Ligase is insensitive to salt carried over from other reaction components (vector or insert DNA) and allows ligation reactions to proceed in alternative reaction buffers with higher levels of salt (e.g., NEBuffer 3.1). |
| Quick Ligation™ Kit | M2200S/L | 25°C (4-37°C) |
N | ATP | Rapid ligation of dsDNA fragments with sticky or blunt ends. Routine cloning. | Buffer contains PEG-do not heat inactivate or transform via electroporation. Will also ligate nicks containing mismatches and 1 or 2 nt gaps with high efficiency. |
| Blunt/TA Ligase Master Mix | M0367S/L | 25°C (4-37°C) |
N | ATP | Fastest ligation of blunt and single base overhang (T/A) substrates. T/A cloning, Best choice for adaptor ligation in NGS library preparation. | One tube master mix format. Contains a proprietary enhancer that increases ligation yields. Contains PEG-do not heat inactivate or transform via electroporation. Will also ligate nicks containing mismatches and 1 or 2 nt gaps with high efficiency. |
| Instant Sticky End Ligase Master Mix | M0370S/L | 25°C (4-37°C) |
N | ATP | Fastest ligation of dsDNA with sticky ends. Routine cloning. | One tube master mix format. Contains a proprietary enhancer that increases ligation yields. Contains PEG-do not heat inactivate or transform via electroporation. Will also ligate nicks containing mismatches and 1 or 2 nt gaps with high efficiency. |
| Electro Ligase® | M0369S | 25°C (4-37°C) |
Y (65°C) |
ATP | Efficient Ligation of nicks, sticky ends, and blunt or T/A ends when a PEG-free mixture is required. Suitable for direct transformation via electroporation. | Contains a proprietary enhancer that increases ligation yields. See also notes for T4 DNA Ligase. |
| T3 DNA Ligase | M0317S/L | 25°C (4-37°C) |
N | ATP | A salt-tolerant enzyme for the ligation of nicks, cohesive ends, and blunt ends in dsDNA. Routine cloning. | Buffer contains PEG-do not heat inactivate or transform via electroporation. Can be used with the PEG-free T4 DNA Ligase Buffer (B0202) with reduced activity. Can also ligate the 3' end of RNA to 5'-pRNA when annealed to a DNA complement. |
| T7 DNA Ligase | M0318S/L | 25°C (4-37°C) |
N | ATP | Allows selective ligation of nicks and cohesive ends in dsDNA, not ligating blunt ends or single-base overhangs. | Buffer contains PEG-do not heat inactivate or transform via electroporation. High specificity for correctly base-paired nicks if used with T4 DNA Ligase Buffer (B0202) instead of supplied T7 DNA Ligase Buffer. |
| E. coli DNA Ligase | M0205S/L | 25°C (4-37°C) |
Y (65°C) |
NAD | Selective ligation of nicks in dsDNA without significant joining of dsDNA fragments regardless of end type. cDNA synthesis. | Used in some cDNA library preparation protocols. |
| HiFi Taq DNA Ligase |
M0647S | 60°C (37-75°C) |
N | NAD | Thermostable NAD dependent ligase that ligates only nicks in dsDNA with the highest discrimination against mismatched bases. Useful for SNP detection through LCR, LDR and other ligation based detection methods. |
Use Thermostable Ligase Reaction Temperature Calculator to estimate incubation temperature. |
| 9°N™ DNA Ligase | M0238S/L | 60°C (45-70°C) |
N | ATP | Thermostable ATP dependent ligase that ligates only nicks in dsDNA with a high discrimination against mismatched bases. Useful for LDR and other ligation based detection methods. | |
| Taq DNA Ligase | M0208S/L | 60°C (37-75°C) |
N | NAD | Thermostable NAD dependent ligase that ligates only nicks in dsDNA with a high discrimination against mismatched bases. Useful for LDR and other ligation based detection methods. | Used in Gibson Assembly® methods. |
| SplintR® Ligase | M0375S/L | 20°C (4-37°C) |
Y (65°C) |
ATP | Ligation of the 3' end of DNA to a 5'-pDNA annealed to an RNA complement. Specific detection of RNA sequences through the ligation of complementary DNA probes. | Ligates fully DNA substrates with high efficiency. Can also ligate the 3' end of RNA to 5'-pDNA when annealed to either DNA or RNA complements. |
| Thermostable 5' AppDNA/RNA Ligase | M0319S/L | 65°C (45-70°C) |
N | N/A | Ligation of the 3-end of ssRNA or ssDNA to a 5'-adenylated ssDNA or 5'-adenylated ssRNA. | DNA to AppDNA ligation occurs with reduced yields. |
| 5'-DNA Adenylation Kit | E2610S/L | 65°C (45-70°C) |
N | ATP | Conversion of 5'-phosphates to 5'-adenylates on ssDNA and ssRNA. | |
| T4 RNA Ligase | M0204S/L | 37°C (4-37°C) |
Y (65°C) |
ATP | ssRNA/ssRNA ligation and ssRNA circularization | |
| T4 RNA Ligase 2 | M0239S/L | 37°C (4-37°C) |
N | ATP | Ligation of dsRNA nicks and ssRNA to ssRNA | Can also ligate the 3' end of RNA or DNA to a 5'-pDNA when annealed to an RNA complement, and the 3' end of RNA to a 5'-pRNA when annealed to an DNA complement, with reduced efficiency. |
| T4 RNA Ligase 2, Truncated | M0242S/L | 25°C (4-37°C) |
Y (65°C) |
N/A | Ligation of the 3'-end of ssRNA to 5' adenylated ssDNA and 5' adenylated ssRNA | |
| T4 RNA Ligase 2, Truncated K227Q | M0351S/L | 25°C (4-37°C) |
Y (65°C) |
N/A | Ligation of the 3'-end of ssRNA to 5' adenylated ssDNA and 5' adenylated ssRNA | Reduction of side products vs M0242. |
| T4 RNA Ligase 2, Truncated KQ | M0373S/L | 25°C (4-37°C) |
Y (65°C) |
N/A | Ligation of the 3'-end of ssRNA to 5' adenylated ssDNA and 5' adenylated ssRNA | Preferred choice for ligation of ssRNA to preadenylated adapters. Reduction of side products vs M0242. |
| RtcB Ligase | M0458S | 37°C |
Y (65°C) |
GTP | Ligation of ssRNA with 3’phosphate or 2’,3’- cyclic phosphate to the 5’- OH of ssRNA. |
