The host strain used for a specific template preparation can impact template quality. JM101 is not recommended. If use of phenol or chloroform can not be avoided an additional ethanol precipitation is recommended. D N A Sequencing Inadequate template preparation is the most common cause of sequencing problems. Qiagen DNA isolation products and alkaline lysis with PEG precipitation have proven to be the most reliable.
Protocols are available at the lab.
Bio 101 GeneKleen Promega Wizard BioRad Prep A Gene 4 Frequent reasons for failure to get good data Failure results when there is an insufficient level of fluorescent termination products for the computer software to assign a sequence. Too little template results in reactions with little or no signal and poor or no base calling. Too much DNA producing reactions which terminate prematurely, often with fewer than 250 bases of reliable sequence data. Template DNA must be free of residual ethanol and salt. Insufficient primer concentration or poor quality.