Cycle Sequencing: Dideoxy-mediated Sequencing Reactions Using PCR and End-labeled Primers.

نویسندگان

  • Joseph Sambrook
  • David W Russell
چکیده

MATERIALS 5x Cycle-sequencing buffer AmpliTaq CS or other exonuclease-deficient versions of Taq DNA polymerase (5 units/μl) EDTA (0.05 M, pH 8.0) Formamide loading buffer Oligonucleotide Primers, radiolabeled at the 5' terminus with 33P or 32P To prepare end-labeled primers, please see Phosphorylating the 5' Termini of Oligonucleotides. Template DNAs Plasmids, cosmids, bacteriophage , and bacteriophage M13 DNAs, purified from largeor small-scale cultures can be used as templates. The table below shows the amounts of each type of template required.

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عنوان ژورنال:
  • CSH protocols

دوره 2006 1  شماره 

صفحات  -

تاریخ انتشار 2006