T4 RNA Ligase 1 (ssRNA Ligase)

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
M0204S     -20       T4 RNA Ligase 1 (ssRNA Ligase) M0204SVIAL -20 1 x 0.1 ml 10,000 units/ml   T4 RNA Ligase Reaction Buffer B0216SVIAL -20 1 x 1.5 ml 10 X   Adenosine-5'-Triphosphate (ATP) B0756AVIAL -20 1 x 0.2 ml 10 mM   50% PEG 8000 B1004SVIAL -20 1 x 1 ml 1 X
M0204L     -20       T4 RNA Ligase 1 (ssRNA Ligase) M0204LVIAL -20 1 x 0.5 ml 10,000 units/ml   T4 RNA Ligase Reaction Buffer B0216SVIAL -20 1 x 1.5 ml 10 X   50% PEG 8000 B1004SVIAL -20 2 x 1 ml 1 X   Adenosine 5'-Triphosphate (ATP) P0756SVIAL -20 1 x 1 ml 10 mM

Application Features

• Labeling of 3'-termini of RNA with 5'-[³²P] pCp (3)
• Inter- and intramolecular joining of RNA and DNA molecules (4,5)
• Synthesis of single-stranded oligodeoxyribo-nucleotides (6)
• Incorporation of unnatural amino acids into proteins (7)
• Ligation of ss-RNA and DNA

Companion Products

• T4 RNA Ligase 1 (ssRNA Ligase), High Concentration
• T4 RNA Ligase 2 (dsRNA Ligase)
• T4 RNA Ligase 2, truncated
• T4 RNA Ligase 2, truncated K227Q
• RNase Inhibitor, Murine

Materials Sold Separately

• T4 RNA Ligase Reaction Buffer 
• Adenosine 5'-Triphosphate (ATP)

1. Addition of DMSO to 10% (v/v) is required for pCp ligation (3).

1. England, T., Gumport, R. and Uhlenbeck, O. (1977). Proc. Natl. Acad. Sci. USA. 74, 4839-4842.
2. Rand, K.N. and Gait, M.J. (1984). EMBO J.. 3, 397-402.
3. England, T. and Uhlenbeck, O. (1978). Nature. 275, 560-562.
4. Romaniuk, P. and Uhlenbeck, O. (1983). R. Wu, L. Grossman and K. Moldave(Ed.), Methods Enzymol.. 100, 52-56. New York: Academic Press.
5. Moore, M.J. and Sharp, P.A. (1992). Science. 256, 992-997.
6. Tessier, D.C., Brousseau, R., and Vernet, T. (1986). Anal. Biochem.. 158, 171-178.
7. Noren, C.J. et al. (1989). Science. 244, 182-188.
8. Silber, R., Malathi, B.G. and Hurwitz, J. (1972). Proc. Natl. Acad. Sci. USA. 69, 3009-3013.

1. 3´ End Labeling of RNA using T4 RNA Ligase 1
2. RNA Circularization using T4 RNA Ligase 1 (ssRNA Ligase) (NEB #M0204)
3. Protocol Ligation of an oligo to the 3’end of RNA using T4 RNA Ligase 1 (NEB #M0204)

• Properties of DNA and RNA Ligases
• RNA Ligase Selection Chart

1. What is the optimal reaction temperature and time for T4 RNA Ligase I?
2. Can T4 RNA Ligase 1 be used to end label single stranded DNA or RNA?
3. Will T4 RNA Ligase 1 ligate double stranded DNA?
4. What is the most common cause of ligation failure when using T4 RNA Ligase 1?
5. Why doesn’t the new T4 RNA Ligase reaction buffer contain ATP?
6. Will the ligation reaction using T4 RNA Ligase 1 produce circles or duplex products?
7. Can T4 RNA Ligase 1 be used to make single stranded RNA/DNA hybrids?
8. Can T4 RNA Ligase 1 ligate triphosphates?
9. Can T4 RNA Ligase 1 be used to add tails to linear duplex DNA?
10. Can a dideoxy be used to block ligation using T4 RNA Ligase 1?
11. Can T4 RNA Ligase 1 be used to incorporate unnatural amino acids into proteins?
12. Will PEG improve ligation with T4 RNA Ligase 1?
13. Will T4 RNA Ligase 1 ligate single stranded DNA?
14. Is PEG 8000 available for purchase?
15. What is the salt tolerance of this RNA ligase?

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

• M0204S_L_v2

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

• M0204S_L_v1_0651802
• M0204S_L_v1_0671804
• M0204S_v1_10008843
• M0204S_v1_10015225
• M0204S_v1_10020007
• M0204L_v1_10032093
• M0204L_v1_10035070
• M0204S_v1_10035072
• M0204L_v1_10048648
• M0204S_v1_10048650
• M0204S_v1_10059648
• M0204S_v1_10062826
• M0204L_v1_10062828
• M0204L_v1_10075416
• M0204S_v1_10075415
• M0204L_v1_10084254
• M0204S_v1_10084255
• M0204L_v1_10096710
• M0204L_v1_10097598
• M0204S_v1_10097601
• M0204L_v1_10103416
• M0204S_v1_10103420
• M0204S_v1_10111889
• M0204L_v1_10111599
• M0204L_v1_10120560
• M0204S_v1_10129005
• M0204L_v1_10129006
• M0204S_v1_10160302
• M0204L_v1_10176560
• M0204S_v1_10184620
• M0204S_v1_10208178
• M0204L_v1_10211008
• M0204S_v1_10221738
• M0204S_v1_10230820
• M0204L_v1_10241174
• M0204S_v1_10249342
• M0204S_v2_10268289
• M0204L_v2_10270368
• M0204S_v2_10289838
• M0204L_v2_10292877
• M0204S_v2_10302398
• M0204L_v2_10308203
• M0204S_v2_10314362
• M0204S_v2_10329188
• M0204S_v2_10327231
• M0204L_v2_10326333

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• T4 RNA Ligase 1 (ssRNA Ligase)
• T4 RNA Ligase Reaction Buffer
• PEG 8000
• Adenosine 5'-Triphosphate (ATP)

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