Mesoplasma florum Assembly

• Methodology Overview
• Assembly Data
  • Genomic Libraries

Methodology Overview

The Mesoplasma florum genome was sequenced using the Whole Genome Shotgun methodology, whereby:

1. High-molecular weight genomic DNA is isolated from Mesoplasma florum L1 (ATCC 33453) and subsequently sheared into smaller-sized fragments (~2 kb, ~4 kb, ~6 kb, ~8 kb, ~10 kb, and ~40 kb).
2. DNA fragments are size-fractionated, ligated into the appropriate vector and subsequently amplified in E. coli cells.
3. The two ends of each cloned DNA fragment are sequenced, creating paired reads.
4. The assembly process uses the paired reads to identify contiguous stretches of sequence (contigs).
5. Contigs are ordered and linked together into larger supercontigs by using paired reads lying in different contigs.
6. Standard finishing methods are applied to resolve low-quality regions (sequence ambiguities) and to obtain additional sequence from specific genome regions that are missing from the original assembly.

   Assembly Data

   Final Assembly

   Sequencing Facts

   • Greater than 10X sequencing coverage of the genome
   • Circular genome with a size of 793,224 bp
   • G+C content 27.02%

   Genomic Libraries: Details

   Sequence reads were derived from random small and large insert plasmid libraries and from a random Fosmid library.

   Library	Type	Vector	Average Insert Size (Kb)	# Attempted Reads	# Reads in Assembly (version 1)
   1	Plasmid	pOT	2.0	11,138	10,155
   2	Plasmid	pOT	4.0	35,136	&emdash;
   3	Plasmid	pJAN	6.0	7,200	6,502
   4	Plasmid	pJAN	8.0	7,392	5,974
   5	Plasmid	pJAN	10.0	7,680	&emdash;
   6	Fosmid	pEpiFOS-5	40.0	3,456	&emdash;