The information about an entry that can not be described using FEATURES or the other fields. For instance, if submitter has the other affiliation to REFERENCE 1, it can be described on COMMENT line.
example COMMENT Human cDNA sequencing project.
To describe sample information in detail, COMMENT lines structured with sets of [item-name] and [item-value] can be used. This strategy is recommended to be used for submissions related to biodiversity and complete genome scale submissions.
example COMMENT ##Genome-Assembly-Data-START## Finishing Goal :: Finished Current Finishing Status :: High Quality Draft Assembly Method :: Newbler v. 2.3 Genome Coverage :: 30x Sequencing Technology :: 454/Illumina ##Genome-Assembly-Data-END##
For MGA submissions. the process for obtaining the submitted sequence data e.g.; (methods for preparing sequences from tissues or cells and processing the sequences for submission) is described.
example COMMENT The CAGE (cap analysis gene expression) is based on preparation and sequencing of concatamers of DNA tags deriving from the initial 20/21 nucleotides from 5' end mRNAs. Full-length cDNAs were at first selected with the Cap-Trapper method. Then, a specific linker (Linker1, some linker contain 5 bp sequences that have 15 variations for each rna sample) containing the ClassIIs restriction enzyme site MmeI was then ligated to the single-strand cDNA and then the second strand of cDNA synthesized. The resulting double-stranded cDNA was cleaved by the restriction enzyme MmeI and a second linker (Linker2) was ligated to the 2 bp overhang at the MmeI cleaved site, to produce a 5' 20/21 tag having two linkers at both sides. The ligation products were separated from unmodified DNA with magnetic beads. The 5' end cDNA tags were released from the beads, and the DNA fragments were amplified in a PCR step by using the two linker-specific primers (Primer1 (uni-PCR), Primer2 (MmeI-PCR)). The desired 32-37 bp tags were purified and ligated to form concatamers, and then the concatamer were fractionated and ligated to the plasmid ZErO-2. The ligations were finally electroporated into DH10b cells (Invitrogen) and obtained plasmids were sequenced with forward primers. CAGE libraries were sequenced with forward primers essentially as described with minor modifications to use zeocin for selection of recombinants. We used in-house developed algorithms for the extraction of tags and for masking the vectors. CAGE tags were extracted with the following parameters: vector masking, minimum 12 bp recognition allowed; linker (13 bp) masking: maximum mismatch, 2 bp allowed; XmaJI site maximum mismatch, 2 bp allowed; tag length, 17-24 bp. Linker1: "Upper oligonucleotide GN6": biotin-agagagagacctcgagtaactataacggtcctaaggtagcgacctagg (5 bp) tccgacGNNNNN and "Upper oligonucleotide N6":