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PCR
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Direct PCR: MightyAmp
> 񃧯-Ver.2
ÃßÃâ Á¶ÀÛÀÌ ÇÊ¿ä¾ø´Â direct PCRÀÌ °¡´É!!
񃧯-Ver.2
Á¦Á¶»ç
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Takara
R071A
MightyAmp¢â DNA Polymerase Ver.2
250 U
(200 ȸ / 50 ¥ìl ¹ÝÀÀ ½Ã)
216,800¿ø
271,000¿ø
09.02 ~ 10.31
Takara
R071B
MightyAmp¢â DNA Polymerase Ver.2
1000 U(R071A x 4)
980,000¿ø
Product index
Overview
PCR FAQs - ÃßõÇÏ´Â PC..
PCR FAQs - Optimizing ..
High Fidelity È¿¼Ò-Pri..
PCR¡¢LA PCR¢â¡¢Hot Sta..
[Cancer Research] HLA ..
½ÇÇ躰 PCR È¿¼Ò ¼±ÅÃÇÏ..
High fidelity PCR: PrimeST..
Á¤È®ÇÑ GXL DNA Polymer..
GXL Premix ŸÀÔ
ÃÖ°í Á¤È®µµ-Max v2.
ÃÖ°í Á¤È®µµ-Max DNA Po..
񃧯-HS DNA Polymerase
HS (Premix)
HS with GC Buffer
Pyrobest DNA Polymeras..
GXL Fast Premix, dye p..
Standard PCR: Ex, LA, rTaq..
TaKaRa Ex Premier¢â DN..
Ex Taq Hot Start Versi..
TaKaRa Ex Taq
LA Taq Hot Start Versi..
LA Taq
Taq Hot Start Version
Recombinant Taq
µ¿°á°ÇÁ¶ ºÐÁÖÇüÅÂ-Taq ..
SpeedSTAR HS È¿¼Ò
Fast PCR¿ë Z-Taq
Taq Antibody
Premix ½Ã¾à (Ex, LA Ta..
Premix ½Ã¾à (Hot Start..
¾÷±×·¹À̵å Taq HS Prem..
Direct PCR: MightyAmp
°ÇÑ ÁõÆø´É·Â- Ver.3
Genotyping¿ë ÃßÃâ½Ã¾à..
񃧯-Ver.2
Dye°¡ Æ÷ÇÔµÈ premix
Colony PCR¿ë premix
Hot Start Version prem..
Fast PCR¿ë premix
PCR¿ë Premix ½Ã¸®Áî
Ư¼ö PCR (Epigenetics, UNG..
EpiTaq HS (for bisulfi..
Multiplex PCR Kit Ver...
¼÷ÁÖÀ¯·¡ DNA ¾ïÁ¦-Taq ..
BcaBEST¢ç DNA Polymera..
TaKaRa Taq¢â HS PCR Ki..
Uracil DNA Glycosylase..
PCR Enzyme (Clontech)
Clontech PCR Enzyme ¼±..
[°³¿ä] So Easy Product
[Cancer Research] Canc..
Advantage 2 Polymerase..
Advantage GC 2 Polymer..
Advantage Genomics LA ..
Advantage HD Polymeras..
Advantage HF 2 PCR Kit
CloneAmp HiFi PCR Prem..
High Fidelity PCR EcoD..
High Yield PCR EcoDry ..
SeqAmp¢â DNA Polymeras..
SuperPlex¢â Premix
Terra PCR Direct Polym..
TITANIUM¢ç Taq DNA Pol..
DNA PCR °ü·Ã Á¦Ç°
Diversify PCR Random M..
LA PCR¢â ¿ë genome DNA..
TaKaRa LA PCR¢â Kit Ve..
TaKaRa PCR Amplificati..
RT-PCR °ü·Ã Á¦Ç°
PrimeScript¢â II High ..
RT-PCR kit, ²Ä²ÄÈ÷ µû..
BcaBEST¢ç RNA PCR Kit ..
PrimeScript¢â High Fid..
PrimeScript¢â One Step..
PrimeScript¢â RT-PCR K..
SMARTer¢ç RACE 5'/3' K..
TaKaRa One Step RNA PC..
TaKaRa RNA LA PCR¢â Ki..
TaKaRa RNA PCR¢â Kit (..
Mutagenesis
Diversify PCR Random M..
PCR Subtraction °ü·Ã Á¦Ç°
Clontech PCR-Select Ba..
Clontech PCR-Select cD..
Clontech PCR-Select Di..
PCR ÀåÄ¡
PCR °ü·Ã ¼Ò¸ðÇ°
TaKaRa PCR Thermal Cyc..
TaKaRa PCR Thermal Cyc..
PCR Accessory Products
Advantage Ultrapure PC..
TaqStart Antibody
Whole Genome SNP Detection
TITANIUM DNA AMPLIFICA..
º´¿øü¤ýÀ¯ÀüÀÚ °ËÃâ (PCR)
¸Åµ¶¿ø¼º ´ëÀå±Õ °ËÃâ¿ë..
Àå°ü ÃâÇ÷¼º ´ëÀå±Õ ¿ë ..
Àå¿° Vibrio °ËÃâ¿ë Pri..
ÄÝ·¹¶ó µ¶¼Ò °ËÃâ¿ë Pri..
Ư¼ö ¼¼±Õ °ËÃâ¿ë Posit..
Ȳ»öÆ÷µµ»ó±¸±Õ °ËÃâ¿ë ..
Bacterial 16S rDNA PCR..
Bacterial 16S rDNA PCR..
Botulinus±Õ °ËÃâ¿ë Pri..
Fungal rDNA (ITS1) PCR..
O-157(º£·Î µ¶¼Ò À¯ÀüÀÚ..
Salmonella±Õ °ËÃâ¿ë Pr..
Shigella ¹× Àå°üħÀÔ¼º..
TaKaRa źÀú±Õ PCR Dete..
TaKaRa PCR Mycoplasma ..
Welchii±Õ °ËÃâ¿ë Prime..
Á¦Ç°¼³¸í
MightyAmp DNA Polymerase´Â ƯÈ÷ 2 kbÁ¤µµ±îÁöÀÇ ÂªÀº ´ÜÆíÀ» ÁõÆøÇϴµ¥ ÀÖ¾î¼ ³ôÀº ¹ÝÀÀ¼ºÀ» °¡Áö´Â ½Å±Ô DNA PolymeraseÀÌ´Ù. º» È¿¼ÒÀÇ °·ÂÇÑ ÁõÆø ´É·Â¿¡ ÀÇÇØ ÀϹÝÀûÀÎ PCR È¿¼Ò¿¡¼ ÁõÆøÀÌ °ï¶õÇÑ PCR ÀúÇØ ¹°ÁúÀ» ¸¹ÀÌ Æ÷ÇÔÇÏ´Â crude »ýü ÃßÃâ¾×À¸·ÎºÎÅÍ PCR ÁõÆøÀÌ °¡´ÉÇÏ´Ù.
MightyAmp DNA Polymerase Ver. 2 ¿¡¼´Â ±âÁ¸ÀÇ MightyAmp DNA Polymerase¿¡ °³·®µÈ buffer¸¦ Á¶ÇÕÇÏ¿© Ç÷¾×, µ¿½Ä¹° Á¶Á÷ µîÀÇ »ýü ½Ã·á¸¦ Á÷Á¢ ¹ÝÀÀ¾×¿¡ ÷°¡ÇÏ´Â ¡¸Direct PCR¡¹ÀÌ °¡´ÉÇÏ°Ô µÇ¾ú´Ù. ±âÁ¸ÀÇ MightyAmp DNA Polymerase¿Í °°ÀÌ PCRÀúÇØ ¹°ÁúÀ» ¸¹ÀÌ Æ÷ÇÔÇÑ crude »ýü ÃßÃâ¾×À¸·ÎºÎÅÍ PCR ÁõÆø»Ó¸¸ ¾Æ´Ï¶ó GC rich, AT rich ¼¿¿¡ °ü°è¾øÀÌ, ±¤¹üÀ§ÇÑ Å¸°ÙÀÇ ÁõÆøÀÌ °¡´ÉÇÏ°Ô µÇ¾ú´Ù. ¼Ò·®ÀÇ ÁÖÇüÀ¸·ÎºÎÅÍÀÇ ÁõÆø¿¡¼µµ ¾çÈ£ÇÑ ¹ÝÀÀ¼ºÀ» ³ªÅ¸³½´Ù.
¶ÇÇÑ 98 ¡É±îÁö polymerase È°¼ºÀ» ¾ïÁ¦ÇÏ´Â °·ÂÇÑ monoclonal Ç×ü¸¦ ÀÌ¿ëÇÏ¿© hot start PCRÀÌ °¡´ÉÇÏ´Ù.
³»¿ë (200 ȸ
*1
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MightyAmp
¢ç
DNA Polymerase Ver.2
200 §¡
2¡¿ MightyAmp Buffer Ver. 2 (Mg
2+
, dNTP plus)
*2
1 §¢ x 5
*1
50§¡ PCR ¹ÝÀÀ½Ã ¿ë·®
*2
4mM Mg
2+
, 800 ¥ìM dNTP Æ÷ÇÔ
º¸Á¸ -20¡É
ÀϹÝÀûÀÎ PCR ¹ÝÀÀ¾× Á¶¼º (total 50 §¡)
»ç¿ë·®
ÃÖÁ¾ ³óµµ
2¡¿MightyAmp Buffer Ver. 2
25 §¡
1¡¿
Primer 1
15 pmol
0.3 ¥ìM
Primer 2
15 pmol
0.3 ¥ìM
»ýü½Ã·á ¶Ç´Â ÃßÃâ¾×
*1
¡Â5 §¡
*1
MightyAmp DNA Polymerase
1 §¡
1.25 U/50 §¡
D.W
up to 50 §¡
*1 ¾Æ·¡ Ç¥¸¦ ÂüÁ¶
<Á¶Á÷»ùÇà ¹× ÃßÃâ¾× »ç¿ë±âÁØ>
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¡Â5 §¡
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2
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3
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»ýü ½Ã·á°áÁ¡ ÃßÃâ¾×
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EDTA ¶Ç´Â heparinÀ» ó¸®ÇÑ Ç÷¾×À» »ç¿ëÇÑ´Ù.
Citric acid (±¸¿¬»ê)À» ó¸®ÇÑ Ç÷¾×À» ÀÌ¿ëÇÏ¸é ¹ÝÀÀ¼ºÀÌ ÇöÀúÇÏ°Ô ÀúÇϵǹǷÎ, Direct PCR¿¡¼´Â ÃßõÇÏÁö ¾Ê´Â´Ù.
Primer ¼³°è
primer´Â °¡´ÉÇÑ primer ¼³°è ¼ÒÇÁÆ®¿þ¾î¸¦ ÀÌ¿ëÇÏ¿© ÃÖÀûÀÎ ¹è¿À» ¼±ÅÃÇÏ°í, Tm°ª*ÀÌ 60¡É ÀÌ»óÀÌ µÇµµ·Ï ¼³°èÇÏ´Â °ÍÀ» ÃßõÇÑ´Ù.
*Tm°ª (¡É)=[(nA+nT)¡¿2]+ [(nC+nG)¡¿4]-5
MightyAmp DNA PolymeraseÀÇ °æ¿ì À̳ë½Å (inosinic acid)À» Æ÷ÇÔÇÏ´ÂprimerÀÇ »ç¿ëÀº ÇÇÇÑ´Ù.
PCR ÁõÆøÁ¶°Ç
Extension ¿Âµµ 68¡É¿¡ 3 step PCRÀÌ Ç¥ÁØÁ¶°ÇÀÌ°í, ÁõÆø±æÀÌ°¡ 2 kbÀÌ»óÀ̰ųª GC rich ¼¿À» ÁõÆøÇÏ´Â °æ¿ì´Â 2 step PCRÀ» ±ÇÀåÇÑ´Ù. [3 Step PCR]
98 ¡É
2 min.
* 1
¡é
98 ¡É
10 sec.
¦¤
60 ¡É
15 sec.
¦¢ 30~40 cycles
68 ¡É
* 2
1 min. / kb
¦¥
[2 Step PCR]
98 ¡É
2 min.
* 1
¡é
98 ¡É
10 sec.
£Ý30~40 cycles
68 ¡É
1 min. / kb
*1
°·ÂÇÑ Hot start Antibody¸¦ Æ÷ÇÔÇϹǷÎ, 98¡É, 2ºÐÀÇ pre-denaturation°úÁ¤ÀÌ ÇʼöÀÓ.
*2
3 step PCRÀÇ °æ¿ìµµ extension¿Âµµ´Â 68¡É·Î ¼³Á¤ÇØ ÁֽʽÿÀ.
PCR»ê¹°
MightyAmp DNA Polymerase¸¦ ÀÌ¿ëÇؼ ÁõÆøÇÑ PCR»ê¹°ÀÇ ´ëºÎºÐÀº 3¡¯ ¸»´Ü¿¡ A°¡ 1¿°±â ºÎ°¡µÇ¾î ÀÖ´Ù.
°ü·ÃÁ¦Ç°
- ¹ü¿ëÀûÀÎ ¿Ã¶ó¿î´õ agarose:
SeaKem LE Agarose
- 5ºÐ¸¸¿¡ Àü±â¿µµ¿ ¿Ï·á! Precast gel system:
FlashGel¢â System
- ÄÄÆÑÆ®ÇÑ 96 well gradient PCR ±â±â:
Clontech PCR Thermal Cycler GP
Keyword : direct, Á÷Á¢
Direct PCR: MightyAmp
°·ÂÇÏ°Ô ÁøÈÇÑ direct PCR¿ë È¿¼Ò
°ÇÑ ÁõÆø´É·Â- Ver.3
DNA °£ÀÌÃßÃâºÎÅÍ genotyping±îÁö ÇϳªÀÇ kit·Î
Genotyping¿ë ÃßÃâ½Ã¾àÆ÷ÇÔ
ÃßÃâ Á¶ÀÛÀÌ ÇÊ¿ä¾ø´Â direct PCRÀÌ °¡´É!!
񃧯-Ver.2