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¸¶Å©·ÎÁ¨ÀÇ SequencingÀº cDNA Library, Shotgun Library µî
Library Á¦ÀÛ¿¡¼ºÎÅÍ Sequencing service¿¡ À̸£´Â
ÀÏ·ÃÀÇ °úÁ¤µéÀ» ½Å¼ÓÇϰí Á¤È®ÇÏ°Ô ¼öÇàÇϰí ÀÖ½À´Ï´Ù.
Bioinformatics ±â¼úÀ» Á¢¸ñÇÑ Blast Search ¹× single contig
ÀÇ ¿Ï¼º¿¡ À̸£±â±îÁöÀÇ ¾÷¹«µéÀ» ¼öÇàÇϸç À̿ܿ¡ Sequencing À» À§ÇØ ÇÊ¿äÇÑ Cell Culture, Colony Picking,
DNA Prepa-ration, PCR ¹×
Purfication, Cell Stock Á¦ÀÛ µî ´Ù¾çÇÑ ºÎ°¡¼ºñ½ºµéµµ ÃÖ»óÀÇ ¼ºñ½º·Î
Á¦°øÇϰí ÀÖ½À´Ï´Ù.
ÀÚÁÖ
¹®ÀÇÇϽô Ç׸ñµéÀ» Á¤¸®ÇÏ¿´½À´Ï´Ù. [FAQ ¹Ù·Î°¡±â] |
Difficult sequencing |
ÀϹÝÀûÀÎ ½ÃÄö½ÌÁ¶°Ç(BigDye v.3.1, AB)¿¡¼´Â ÁÁÀº °á°ú¸¦ ¾òÀ» ¼ö
¾ø´Â template¸¦ ´ë»óÀ¸·Î ÇÏ´Â ½ÃÄö½Ì ¼ºñ½º ÀÔ´Ï´Ù. ÀϹÝÀûÀÎ
½ÃÄö½Ì°ú´Â ´Ù¸¥ Á¶°Ç°ú ½Ã¾àÀ¸·Î ÁøÇàµÇ¸ç, °¡°Ýü°è ¶ÇÇÑ ´Ù¸¨´Ï´Ù.
Á» ´õ ÀÚ¼¼ÇÑ Á¤º¸¸¦ ¿øÇϽøé [¿©±â]¸¦ ´©¸£½Ã±â ¹Ù¶ø´Ï´Ù.
*»ùÇÃÀÇ Æ¯À̼º¿¡ µû¶ó ¼º°øÀûÀÎ °á°ú¸¦ ¾òÁö ¸øÇÏ´Â °æ¿ìµµ
ÀÖ½À´Ï´Ù. |
16s rRNA |
Bacterial cellÀ» ÀÌ¿ëÇÏ¿© ÇÙ»êÃßÃâ->PCR->Sequencing->AssembleµîÀÇ
¸ðµç °úÁ¤À» ¼öÇàÇØ µå¸®´Â ¼ºñ½º ÀÔ´Ï´Ù.
domain BacteriaÀÇ °æ¿ì 1,300base pairsÀÌ»óÀÇ
°á°ú¸¦ º¸ÀåÇÏ¿© µå¸³´Ï´Ù.
»ùÇà Áغñ´Â [¿©±â]¸¦ Ŭ¸¯ÇÏ¿© Âü°íÇØ Áֽñ⠹ٶø´Ï´Ù. |
18s rRNA(ITS region) |
»ý¸íüÀÇ Á¾À» µ¿Á¤ÇÏ´Â ¹æ¹ý Áß Çϳª·Î ¿°±â¼¿ Á¤º¸¸¦ ÀÌ¿ëÇÏ´Â ¹æ¹ýÀÌ ³Î¸® ÀÌ¿ëµÇ°í
ÀÖ½À´Ï´Ù. ¿°±â¼¿ Á¤º¸´Â small subunit ribosome À» ±¸¼ºÇÏ´Â rRNA¸¦ ¿°±â ¼¿ ºÐ¼®ÇÏ¿© ÀÌ¿ëÇÕ´Ï´Ù.
ÁøÇÙ»ý¹°ÀÇ °æ¿ì 18S(ITS 5.8s) rRNA full sequencingÀ» ÅëÇØ ¿°±â¼¿À» ºÐ¼®ÇÏ°í ±× °á°ú¸¦
rRNA database(NCBI)¿Í ºñ±³ÇÏ¿© ´ë»ó ¹Ì»ý¹°ÀÌ ¾î¶² ¹Ì»ý¹°°ú À¯»çÇÑÁö¸¦ È®ÀÎÇÏ´Â ¼ºñ½º ÀÔ´Ï´Ù.
»ùÇà Áغñ´Â [¿©±â]¸¦ Ŭ¸¯ÇÏ¿© Âü°íÇØ Áֽñ⠹ٶø´Ï´Ù.
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ITS+26S(D1/D2 region)  |
»ý¸íüÀÇ Á¾À» µ¿Á¤ÇÏ´Â ¹æ¹ý Áß Çϳª·Î ¿°±â¼¿ Á¤º¸¸¦ ÀÌ¿ëÇÏ´Â ¹æ¹ýÀÌ ³Î¸® ÀÌ¿ëµÇ°í
ÀÖ½À´Ï´Ù. ¿°±â¼¿ Á¤º¸´Â Ribosomal RNA À¯ÀüÀÚ¸¦ ¿°±â¼¿ ºÐ¼®ÇÏ¿© ÀÌ¿ëÇÕ´Ï´Ù.
ÁøÇÙ»ý¹°ÀÇ °æ¿ì ITS region ¹× 26S(D1/D2) region sequencing À» ÅëÇØ ¿°±â¼¿À» ºÐ¼®ÇÏ°í ±× °á°ú¸¦ rRNA
database(NCBI)¿Í ºñ±³ÇÏ¿© ´ë»ó ¹Ì»ý¹°ÀÌ ¾î¶² ¹Ì»ý¹°°ú À¯»çÇÑÁö¸¦ È®ÀÎÇÏ´Â ¼ºñ½º ÀÔ´Ï´Ù.
»ùÇà Áغñ´Â [¿©±â]¸¦ Ŭ¸¯ÇÏ¿© Âü°íÇØ Áֽñ⠹ٶø´Ï´Ù. |
Primer walking |
single primer extensionÀ¸·Î Çѹø¿¡ Àо ¼ö ¾ø´Â plasmid
³ª PCR productÀÇ ¿°±â¼¿À» ºÐ¼®ÇÏ´Â ¼ºñ½º À̸ç, º¸Åë 2-10kb Á¤µµÀÇ ½ÃÄö½º Á¤º¸¸¦ ¾ò°íÀÚ ÇÒ¶§ ÀÌ¿ëÇÏ´Â
¹æ¹ýÀÔ´Ï´Ù.°í°´´Ô²²¼ Á¦°øÇϽðųª ÁöÁ¤ÇØÁֽŠprimer·Î end sequencingÀ» ¼öÇàÇÑ ÈÄ ¾òÀº °á°ú¹°·ÎºÎÅÍ
internal primer¸¦ µðÀÚÀÎÇÏ¿© Á¦ÀÛÇÕ´Ï´Ù. »õ·Î¿î Primer·Î °°Àº template¸¦ ¹ÝÀÀÇÏ¿© ¾òÀº °á°ú·ÎºÎÅÍ
¶Ç ´Ù½Ã ÀûÁ¤ÇÑ À§Ä¡¿¡¼ internal primer¸¦ µðÀÚÀÎÇÏ¿© °è¼Ó ¿¬ÀåÇÕ´Ï´Ù. 1ȸ walking ¼öÇà¿¡ 4ÀÏ
Á¤µµ°¡ ¼Ò¿äµÇ¸ç, ÇÑ ¹æÇâ ´ç ¾à 500bp~800bp ¿¬ÀåÀÌ °¡´ÉÇÕ´Ï´Ù.
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Cloning |
PCR ProductÀÇ ContaminationÀÇ °æ¿ì ¿©·¯ Á¾·ùÀÇ Product°¡
Á¸ÀçÇÏ¿© sequencing °á°ú°¡ ÁÁÁö ¾ÊÀº °æ¿ì(sequence PeakÀÇ °ãÄ¡´Â Çö»ó) CloningÀ» ÅëÇØ
°³¼±ÇÒ ¼ö ÀÖ½À´Ï´Ù.
PCR Product¸¦ vector (T-vector or Blunt-vector)¿¡
»ðÀÔÇÏ¿© Plasmid ¸¦ ¸¸µì´Ï´Ù.
Plasmid¸¦ Bacteria cell¿¡ ÁÖÀÔÇÏ¿© ¹è¾çÈÄ
cell extraction ÇÕ´Ï´Ù.
PCR½Ã »ç¿ëÇÏ´Â DNA polymerase(pfu-taq
»ç¿ë À¯¹«)¿¡ µû¶ó vector(T-vector or Blunt-vector)¸¦
´Ù¸£°Ô »ç¿ëÇÕ´Ï´Ù.
µû¶ó¼ ÁÖ¹® ÀÇ·ÚÀü¿¡ »ç¿ëÇϽŠtaqÀÇ ±â´É À¯¹«¸¦
Ç¥±âÇÏ¿© Áֽñ⠹ٶø´Ï´Ù. (with pfu ¶Ç´Â without pfu)
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Human mtDNA Full Length Sequencing  |
16,659 bpÀÇ human mitochondria DNA À¯ÀüÀÚ¸¦ ÁõÆøÇÏ¿© ¸ðµç ¿°±â¼¿À» ºÐ¼®ÇÏ´Â ¼ºñ½º.
- PCR, sequencing, assemble µî ¸ðµç °úÁ¤ ¼öÇà.
- 15 PCR primer setÀ» ÀÌ¿ëÇÏ¿© PCR product »ý¼º, 40°³ sequencing primer¸¦ ÀÌ¿ëÇÏ¿© ¿°±â¼¿
ºÐ¼®, °¢°¢ÀÇ seqeucning data¸¦ ÀÌ¿ëÇÏ¿© ÇϳªÀÇ full contig »ý¼º.
- ¼Ò¿ä±â°£ : Sample QC È®ÀÎ ÈÄ 14ÀÏ.
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Microsatellite Analysis(VNTRs) Service |
Microsatellite Analysis(VNTRs) service encompasses
a wide variety of genotyping, DNA profiling, and mutation detection
techniques for medical, environmental, and agricultural research.
Macrogen Inc. provides the Microsatellite Analysis(VNTRs) service
based on our accumulated experience and know-how in genomics. In general,
it is used for amplified fragments' size check only. Since PCR
amplification
is not available through this service, fluorescent-labeled PCR products
should be
supplied as Microsatellite Analysis(VNTRs) samples.
Service Applications :
> Microsatellite Instability
> Amplified Fragment Length Polymorphism (AFLP)
Analysis
> Terminal Restriction Fragment Length Polymorphism
(T-RFLP) Analysis
> Relative Fluorescent Quantization - Loss of
Heterozygosity (LOH), Aneuploidy Assays,
and
Large Chromosomal Deletion Detection
> Sequence-related amplified polymorphism (SRAP)
provides a useful tool for estimation of genetic
diversity and phenetic relationships in natural
and domesticated populations.
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PCR optimization |
º¸³»ÁֽŠgenomic DNA¿¡¼ PCR Product¸¦ ¸¸µé¶§ ÃÖÀûÀÇ annealing
TMÀ» ã¾Æ ÃÖÀûÈµÈ PCR Product¸¦ »ý»êÇϵµ·Ï Á¶°ÇÀ» Àâ¾ÆÁÖ´Â ¼ºñ½º ÀÔ´Ï´Ù.
gDNA¿¡¼ ÃÖÀûÀÇ PCR Product°¡ »ý»êµÇ¾î¾ß ³ôÀº ǰÁúÀÇ
sequencing data¸¦ ¹Þ¾Æº¸½Ç ¼ö ÀÖ½À´Ï´Ù. |
SNP |
SNP analysis service is highly flexible to any
specific project requirements.
SNPs (single nucleotide polymorphisms) occurs
every 100 to 300 bases along the human genome
and account for about 90% of all genetic variations. SNPs could predispose
people to disease
or influence their response to a drug.
We offer a complete package of SNP discovery
sequencing services by this step below,
PCR optimization => PCR amplification
=> Purification => Sequencing => SNP
discovery
We provide high-throughput SNP Analysis
by using ABI3730XL, automated DNA sequencer,
and the result will be available within
approximately two (2) weeks. |
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