You will be presented with the full text of a biomedical research paper. Please write an abstract for this paper. Your response should include the abstract and no additional text.

Paper text:
earlier , techniques like liquid chromatography , immunoassay , electrophoresis , and so forth were used in species identification . in recent years
, use of dna has been popularized due to its specificity and stability . among types of dna [ 4 , 5 ] , mitochondrial
mitochondria follow clonal inheritance   as only mother to contributes to mitochondria ; its genome does not undergo recombination ; thus , genetic material will be passed onto the next generation unchanged .
reports also suggest that mitochondrial genome is accumulating high percentage of neutral mutations which is helpful in species identification .
pcr amplification of cytochrome b gene has been used in differentiation of meats from buffalo , emu , and crocodile .
cytochrome b has also been used in differentiation of processed products like canned tuna , vegetable oil , and tomato sauce .
other mitochondrial genes like 12s and 16s rrna are extensively used in mammalian species identification .
numerous molecular techniques have been developed based on the use of mitochondrial 12s rrna gene identification like rapd fingerprinting , dna hybridization , restriction fragment length polymorphism [ 11 , 12 ] , real time pcr , and so forth .
mitochondrial 12s rrna based pcr- rflp has been used to differentiate peacock from other poultry species   and nilgai from cattle and buffalo   as well as differentiation of different deer species belonging to the family cervidae .
mouse deer also known as chevrotain are small ungulates that belong to the family tragulidae found in forests of south and southeast asia .
indian spotted mouse deer moschiola indica has been recently segregated as a species separately from moschiola meminna .
indian mouse deer , the smallest ungulate in world , lives in solitary or pairs , feeds on plant material , and weighs 1.5 to 18  lbs . like other ruminants ,
it is known to evolve from oligocene 34 million years ago and remained as a primitive ruminant .
mouse deer has been considered as schedule i animal in wildlife protection act 1972 as its population is declining due to poaching .
the poaching is for its skin and meat largely for the pot and often meat is sold in local market ( iucn red list threatened species 2013 , http://www.iucnredlist.org/details/136585/0 ) . in the present study , partial sequence of mitochondrial 12s rrna gene in indian mouse deer has been characterized for the first time as a tool for species differentiation using pcr - rflp .
hairs were collected from indian spotted mouse deer under captivity at pilikula zoo , mangalore , karnataka , india , and brought to the laboratory under cold conditions .
genomic dna was isolated from hair follicles using qiamp tissue extraction kit as per the manufacturer 's instructions .
the isolated dna was checked for integrity in 0.8% agarose gel electrophoresis in tris - acetate edta buffer ( 40  mm tris - acetate , 2  mm edta , ph 8.0 ) .
mitochondrial 12s rrna gene was amplified using reported   universal primers of 12s rrna , forward primer : 5 caa act ggg att aga tac ccc act at 3 and reverse primer : 5 gag ggt gac ggg cgg tgt gt 3. reaction was set up in 50  l volume consisting of 10x buffer 5.0  l ( 10  mm tris - hcl , 50  mm kcl , and 1.5  mm mgcl2 ) , 0.2  mm dntps , 20  pmol of each primer , 200  ng template , and 1.5  u proof reading enzyme ( fermentas ) . amplification was carried out in a thermal cycler comprising initial denaturation at 94c for 5  min , followed by 35 cycles of denaturation 94c for 45  sec , annealing at 59c for 45  sec , and extension at 72c for 1.0  min .
the amplicon was separated in 1% agarose gel electrophoresis and the gel spliced product was purified using qiaquick gel extraction kit as per manufacturer 's instructions ( qiagen ) .
the purified product was ligated into pjet 2.1 blunt end cloning vector ( invitrogen ) using t4 dna ligase .
the ligated product was transformed in to e. coli dh5 competent cell as per transformation protocol by chung et al .
( 1989 ) and plated onto lb agar with ampicillin ( 50  g / ml ) as marker .
the obtained colonies were picked into 5  ml lb broth with ampicillin and grown at 37c overnight under constant shaking at 200  rpm .
reaction mixture consisted of 10x buffer o 2  l , plasmid ( 1  g / ml ) 5  l , and not1 2  l and final volume was made to be 20  l .
mixture was incubated at 37c for 16  h and digestion pattern was checked by running 1% agarose gel electrophoresis containing 0.5  g / ml ethidium bromide .
mitochondrial 12s rrna gene sequence of indian mouse deer was analysed with sequences of other deer species like barking deer ( af294731 ) , sambar ( m35875 ) , sika deer ( ay184433 ) , chital ( dq017832 ) , hog deer ( ay775785 ) , and himalayan musk deer ( ay847268 ) available at ncbi genbank using dnastar software .
sequence was also used to construct rflp plot with common restriction enzymes used in other deer species for species identification .
pcr amplification has provided 12s mitochondrial rrna gene of nearly 437  bp ( figure 1 ) long nucleotide sequence which was visualized by gel electrophoresis .
the recombinant plasmid encoding the amplicon was sequenced which also provided 437  bp sequence .
further sequence was analysed in silico with other available 12s mitochondrial rrna sequences deer species with the help of lasergene ( dnastar inc . ,
alignment report at nucleotide level showed 90.04 , 90.08 , 90.04 , 91.2 , 90.04 , and 90.08 percent identity with sika deer , sambar , hog deer , musk deer , chital , and barking deer , respectively .
phylogenetic relationship shows early evolution of mouse deer as separate cluster different from other deer species ( figure 3 ) .
the nucleotide sequence was further analysed with the help of lasergene ( dnastar inc . ,
fragments predicted in software which can not be separated in gel electrophoresis have been ignored .
digestion with rsai , ddei , bsri , and bstsfi is expected to reveal fragments of 360  bp , 348  bp , 179 + 249  bp , and 146 + 291  bp , respectively ( figure 4 and table 1 ) .
many types of molecular markers are used in species identification ; among them mitochondrial 12s and 16s rrna gene have been used extensively due to clonal inheritance of mitochondria without recombination and sequence change . over a period mitochondrial genome is also accumulating neutral mutations , leading to change in restriction enzyme recognition patterns which help in species identification . in the present study
mitochondrial 12s rrna gene was used as molecular marker for differentiation of indian mouse deer from other deer species .
amplicon comprising ~437  bp was amplified using universal primers which can amplify corresponding region of various organisms including birds and insects .
sequencing and blast analysis of the sequence at ncbi confirmed its identity with other deer species .
sequence analysis with mitochondrial 12s rrna gene of other deer species has shown more than 90% similarity which confirms the close relationship with other deer species .
mouse deer belongs to the family tragulidae , known to evolve very early from oligocene as primitive ruminant .
it is also different from other deer as the third chamber of rumen is poorly developed
. from phylogenetic analysis it is evident that it evolved very early as a separate cluster from other deer species family cervidae ( figure 3 ) .
though nucleotide sequence showed more than 90% homology , the difference in remaining sequence can be used in species differentiation with restriction fragment length polymorphism ( rflp ) .
restriction digestion with rsai has given fragment of 360  bp which is similar to musk deer but barking deer ( 175 + 190  bp ) and chital , hog deer , sambar , and sika deer ( 152 + 212  bp ) ( gupta et al . , 2008 )
thus , rsai enzyme can be used to differentiate indian mouse deer from other deer except musk deer ( figure 4 and table 1 ) .
the mitochondrial 12s rrna gene of mouse deer is having two recognition sites for both bsri and bstsfi which is likely to produce two fragments ( 179 + 249  bp and 146 + 291  bp resp .
, ) but musk deer sequence yields only one fragment as it has one recognition site .
this can be used to differentiate mouse deer from musk deer ( figure 4 and table 1 ) .
poaching for the purpose of meat is main threat to indian mouse deer . with the help of predicted pattern in pcr - rflp
, mouse deer can be identified using genomic dna isolated from skin or meat samples thereby providing legal molecular aid in wildlife forensics .
in the present study nearly 437  bp mitochondrial 12s rrna gene of indian mouse deer was amplified .
the amplicon was sequenced and analysed in silico with other available sequences of deer species .
sequence analysis and restriction mapping in lasergene ( dnastar inc . , madison , wi , usa ) confirmed the usefulness of 12s rrna gene as molecular marker for differentiation of mouse deer from other species .
thus , it can be concluded that pcr - rflp based analysis of mitochondrial 12s rrna gene can be used as a tool for identification of mouse deer .
this will be useful in providing molecular aid in wildlife forensics and conservation of the mouse deer .
mitochondrial 12s rrna has proven to be a useful molecular marker for better conservation and management of the endangered species 
 . polymerase chain reaction - restriction fragment length polymorphism ( pcr - rflp ) of the mitochondrial 12s rrna gene has proven to be a reliable and efficient tool for the identification of different indian deer species of family cervidae . in the present study , mitochondrial 12s rrna gene sequence of mouse deer ( moschiola indica ) belonging to the family tragulidae 
 was characterized and analysed in silico for its use in species identification . 
 genomic dna was isolated from the hair follicles and mitochondrial 12s rrna gene was amplified using universal primers . 
 pcr product was cloned and sequenced for the first time . 
 the sequence of mouse deer showed 90.04 , 90.08 , 90.04 , 91.2 , 90.04 , and 90.08% identities with sika deer , sambar , hog deer , musk deer , chital , and barking deer , respectively . 
 restriction mapping in lasergene ( dnastar inc . 
 , madison , wi , usa ) revealed that mouse deer mitochondrial 12s rrna gene sequence can be differentiated from the other deer species in pcr - rflp using rsai , ddei , bsri , and bstsfi . with the help of predicted pattern 
 , mouse deer can be identified using genomic dna from a variety of biomaterials , thereby providing molecular aid in wildlife forensics and conservation of the species .