Cold shock protein compositions and methods and kits for the use thereof
View Patent ↗The present invention provides cold shock protein-containing compositions for improved DNA synthesis reactions with improved reactivity, methods for synthesizing DNA using such compositions, kits for use in such methods, and DNA compositions yielded by such methods. The present invention further provides cold shock protein-containing compositions for the identification of endoribonuclease cleavage sites, methods for identifying endoribonuclease cleavage sites using such compositions, and kits for use in such methods.
1. A composition for DNA synthesis comprising a cold shock protein A (CspA) protein of Escherichia coli ; and a thermostable DNA polymerase, wherein the composition comprises more than 0.5 μg of the CspA protein per 25 μL of the composition.
2. The composition according to claim 1 , further comprising at least one additional DNA polymerase.
3. The composition according to claim 2 , wherein at least one DNA polymerase possesses 3′→5′ exonuclease activity and at least one DNA polymerase possesses substantially no 3′→5′ exonuclease activity.
4. The composition according to claim 1 , further comprising at least one primer and at least one deoxyribonucleotide.
5. The composition according to claim 4 , wherein the deoxyribonucleotide comprises a deoxyribonucleotide triphosphate.
6. The composition according to claim 4 , further comprising a liquid vehicle.
7. The composition according to claim 6 wherein the liquid vehicle comprises a reaction buffer solution.
8. A composition for reverse transcription reaction comprising:
a CspA protein of Escherichia coli ; and
a reverse transcriptase of Moloney Murine Leukemia Virus or Avian Myeloblastosis Virus,
wherein the CspA protein is present in the composition at a concentration ranging from about 0.5 μg to about 20 μg of the CspA protein per 20 μL of the composition.
9. The composition according to claim 8 , further comprising at least one primer and at least one deoxyribonucleotide.
10. The composition of claim 9 , further comprising a liquid vehicle.
11. The composition of claim 10 , wherein the liquid vehicle comprises a reaction buffer solution.
12. The composition of claim 9 , further comprising an oligo (dT) primer or an oligonucleotide primer having a random sequence or a combination thereof.