Fast hybridization for next generation sequencing target enrichment
The present invention relates to compositions and methods of target enrichment or selection of nucleic acids using hybridization, which can be used in, e.g., next-generation sequencing.
1. A composition for nucleic acid hybridization, comprising
a salt of a divalent cation that has a concentration in the range from about 100 mM to about 600 mM,
a buffering agent, wherein said salt and said buffering agent are present in a molar ratio of about 2.5:1 to about 60:1, and
a volume-excluding/thickening agent that (i) is selected from the group consisting of hydroxypropyl methyl cellulose (HPMC), hydroxyethyl methyl cellulose (HEMC), hydroxybutyl methyl cellulose, hydroxypropyl cellulose, methyl cellulose, and hydroxyl methyl cellulose and (ii) has a concentration in the range from about 0.002% to about 0.1% w/w.
2. The composition of claim 1 , wherein the buffering agent has a concentration in the range from about 10 mM to about 40 mM.
3. The composition of claim 1 , wherein the divalent cation is selected from the group consisting of magnesium, calcium, manganese, cobalt, zinc, and cadmium.
4. The composition of claim 1 , wherein the buffering agent is selected from the group consisting of Tris, HEPES, TAPS, Tricine, Bicine, Bis-Tris, NaOH, KOH, TES, EPPS, and MOPS.
5. The composition of claim 1 , wherein the divalent cation is magnesium.
6. The composition of claim 5 , wherein the salt is MgCl 2 , the volume-excluding/thickening agent is HPMC, and the buffering agent is Tris.
7. The composition of claim 6 , wherein said salt has a concentration of about 308 mM, said HPMC has a concentration of about 0.00834%, and said buffering agent has a concentration of about 20 mM.
8. The composition of claim 1 , wherein the pH of the composition is about 7-7.
9. A buffer concentrate for preparing a hybridization solution, wherein dilution of said buffer concentrate by a factor ranging from 2-fold to 20-fold yields the composition of claim 1 .
10. The buffer concentrate of claim 9 , wherein dilution of said buffer concentrate by a factor of 5-fold yields said composition.
11. A kit for nucleic acid hybridization, comprising the buffer concentrate of claim 9 and packaging material therefor.