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. Author manuscript; available in PMC: 2011 Feb 1.
Published in final edited form as: Methods. 2010 Jan 12;51(4):458–463. doi: 10.1016/j.ymeth.2010.01.001
Step Start temp End temp Time
1 −140° C −85° C 1.0 hr
2   −85° C −85° C variable: 24–60 hrs
3   −85° C −20° C 13.0 hrs
4   −20° C −20° C 6.0 hrs