Dabeen Park/Question

1. If there no standard gene, using equipment can make short/long sequence.

   How we make the short/long sequence and how we connect the sequence bioinformatically?

   Is there some specipic position?

 

A : It is not only one answer but it can get some idea.

http://www6.appliedbiosystems.com/support/tutorials/pdf/quant_pcr.pdf

Reference : Creating Standard Curves with Genomic DNA or Plasmid DNA Templates for Use in Quantitative PCR

 

2. When DNA duplication, how we find start point like OriC

   Each okazaki fragment combine later but there are no missing part?

   Like 'ATGGGCCTCCAATG', the first fragment start from last ATG,

   the second fragment start from first ATG, but that is not triple size.

 

A : "After the replication fork has passed by, the lagging strand is left as a series of Okazaki fragments with gaps (i.e., spaces from which one or more nucleotides are missing) between them. In addition, there are short pieces of RNA primer alternating with newly-synthesized DNA. Joining the Okazaki fragments to give a complete strand of DNA is accomplished by two—or perhaps three—enzymes working in succession: Ribonuclease H(RNase H), DNA polymerase I (Pol I), and DNA ligase. Only the last two enzymes are involved in the classical model. DNA polymerase I degrades the RNA primers and fills the resulting gaps. Finally, DNA ligase joins the sugar phosphate backbone (Fig. 10.13). In an alternate model, RNase H, which typically degrades the RNA strand of DNA:RNA double helixes, removes most of each RNA primer and DNA polymerase I only removes the last few bases of the RNA primers."

Reference : David P. Clark, Nanette J. Pazdernik, in Molecular Biology (Second Edition), 2013 Cell Division and DNA Replication