Which methodology is used while sequencing the total DNA from a cell? Explain it in detail.

Chhaya Patil · 18th Nov 2019 .

Rojalin Patra · 18th Nov 2019 .

Sequencing the total DNA from a cell (both coding and non-coding regions) and later assigning the different regions with functions is called sequence annotation. This method involves following steps :

(i) The whole DNA of the cell is isolated and broken randomly into fragments.

(ii) They are inserted into specialised vectors like BAC (bacterial artificial chromosome) and YAC (yeast artificial chromosome).

(iii) The fragments are cloned in suitable hosts like bacteria and yeast. Multiple copies of DNA fragments can be obtained by PCR (polymerase chain reaction).

(iv) The fragments are sequenced as annotated DNA sequences.

(v) The sequences were then arranged on the basis of some overlapping regions. It necessitated the generation of overlapping fragments for sequencing.

(vi) Computer based programmes were used to align the sequences.

(vii) The sequences were then annotated and assigned to different chromosomes.

(viii) With the help of polymorphism in micro- satellites and restriction endonuclease recognition sites, the genetic and physical maps of the genome can be prepared.

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