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Home > Learning center > PCR > ´Ù¾çÇÑ ½ÇÇè Àû¿ë »ç·Ê > ´Ù¾çÇÑ ½ÇÇè Àû¿ë <Multiplex PCR>

´Ù¾çÇÑ ½ÇÇè Àû¿ë <Multiplex PCR>

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Multiplex PCR Àû¿ë ¿¹½Ã: SuperPlex¢â Premix¸¦ »ç¿ëÇÑ ºü¸£°í °£ÆíÇÑ multiplex PCR
PCR¿¡¼­ ¹ÝÀÀ ´Ü°¡ ¶Ç´Â Á¦ÇÑµÈ »ùÇÃÀÇ ¾çÀÌ ¹®Á¦¶ó¸é single PCR ´ë½Å multiplex PCRÀ» »ç¿ëÇÏ¿© ´õ¿í È¿°úÀûÀÎ ½ÇÇèÀ» ÁøÇàÇÒ ¼ö ÀÖ´Ù. ±×·¯³ª multiplex PCR¿¡¼­´Â ÇѹøÀÇ ¹ÝÀÀ¿¡¼­ ´Ù¾çÇÑ Å¸°ÙÀÇ ÀÏÁ¤ÇÑ ÁõÆø È¿À²À» ¾ò±â À§ÇÑ ÃÖÀûÈ­ °úÁ¤ÀÌ ÇÊ¿äÇÏ´Ù´Â ÇÑ°è°¡ ÀÖ´Ù. SuperPlex™ Premix (Code 638543)¸¦ »ç¿ëÇÏ¸é ´ÙÀ½°ú °°ÀÌ multiplex PCRÀ» ´õ¿í °£´ÜÇÏ°Ô ÁøÇàÇÒ ¼ö ÀÖ´Ù.
  • Ÿ»ç Á¦Ç° ´ëºñ ´Ù¾çÇÑ Å¸°ÙÀ» ±ÕÀÏÇÏ°Ô ÁõÆøÇÏ´Â multiplex PCR premix
  • Ÿ»ç Á¦Ç° ´ëºñ ¹ÝÀÀ ½Ã°£ÀÌ ºü¸¥ multiplex PCR premix
  • °£´ÜÇÑ workflow : primer, sample, H2O¸¸ ³ÖÀ¸¸é Áغñ ³¡

¡á Introduction
Multiplex PCRÀ» »ç¿ëÇϸé single PCRº¸´Ù ´õ ¸¹Àº Ÿ°ÙÀÌ ÁõÆøµÇ±â ¶§¹®¿¡, ÀûÀº ½Ã°£°ú ½ÇÇèȽ¼ö·Îµµ ´õ¿í ¸¹Àº Á¤º¸¸¦ ¾òÀ» ¼ö ÀÖ´Ù. ÀÌ·¯ÇÑ ÀÌÀ¯·Î multiplex PCRÀº mutation detectionÀ̳ª, pathogen identification, Áúº´ Áø´Ü¿¬±¸, Àü¿°º´ ½ºÅ©¸®´× µîÀÇ ºÐ¾ß¿¡¼­ ³Î¸® »ç¿ëµÇ°í ÀÖ´Ù.

ºÐÀÚ Áø´ÜºÐ¾ßÀÇ ¿¬±¸¿¡¼­´Â ´Ù¼öÀÇ »ùÇÿ¡¼­ Á¦½Ã°£ ¾È¿¡ ÀÇ¹Ì ÀÖ´Â °á°ú¸¦ ¾ò±â À§ÇØ ½ÇÇèÀÇ ¼Óµµ¿Í È¿À²ÀÌ ¸Å¿ì Áß¿äÇÏ´Ù. ½ÇÇèÀÇ workflow¸¦ °³¼±ÇÏ¸é ½ÇÇè ½Ã°£À» Àý°¨ÇÒ ¼ö ÀÖÀ¸¸ç, ÆÄÀÌÆê »ç¿ë ´Ü°è¸¦ ÃÖ¼ÒÈ­ÇØ ¿À¿°À̳ª ¿¡·¯°¡ ¹ß»ýÇÒ °¡´É¼ºÀ» ÁÙÀÌ°í, ¹Ýº¹ÀÛ¾÷¿¡¼­ »ý±æ ¼ö ÀÖ´Â ºÎ»óÀÇ À§ÇèÀ» ÃÖ¼ÒÈ­ÇÒ ¼ö ÀÖ´Ù. ¶ÇÇÑ ½ÇÇè °úÁ¤À» ´Ü¼øÈ­ÇÏ¸é ¿¬±¸ÀÚµé °£ÀÇ ½ÇÇè Ç¥ÁØÈ­¸¦ °¡´ÉÇÏ°Ô ÇÏ¿© ½ºÅ©¸®´× ÇÁ·ÎÁ§Æ®¸¦ È®ÀåÇÏ´Â °ÍÀÌ ¿ëÀÌÇÏ´Ù.

Multiplex PCRÀº ¸Å¿ì À¯¿ëÇÑ ¹æ¹ýÀÎ ¹Ý¸é ½ÇÇèÀÇ ¼¼ÆÃÀÌ ½±Áö ¾Ê´Ù. ÀÌ ¹æ¹ýÀº ±ä ½Ã°£ÀÇ ÃÖÀûÈ­ °úÁ¤ÀÌ ÇÊ¿äÇÏ°í, ¶Ç´Ù¸¥ °úÁ¦·Î´Â primerÀÇ µðÀÚÀΰú ¸ðµç primer°¡ ÀÛµ¿ÇÏ´Â PCR Á¶°ÇÀ» ã¾Æ¾ßÇÏ´Â Á¡ÀÌ ÀÖ´Ù. ÀÌ·¯ÇÑ °úÁ¤À» ´Ü¼øÈ­ÇÏ°í ½ÇÇèÀÇ workflow¸¦ °£¼ÒÈ­Çϱâ À§ÇØ multiplex PCRÀ» »ç¿ëÀÌ Æí¸®ÇÑ master mix ½Ã¾àÀ¸·Î ÁøÇàÇÒ ¼ö ÀÖ´Â SuperPlex™ Premix (Code 638543)À» °³¹ßÇß´Ù. SuperPlex™ Premix´Â °í¼öÀ² PCR È¿¼ÒÀÎ Titanium Taq DNA polymerase ±â¹ÝÀÇ 2X hot strart PCR master mix·Î multiplex PCR¿¡ ÃÖÀûÈ­ µÇ¾îÀÖ´Ù. ÀÌ premix ½Ã¾à¿¡´Â multiplex PCR¿¡ ÇÊ¿äÇÑ ¸ðµç ½Ã¾àÀÌ Æ÷ÇÔµÇ¾î ¶§¹®¿¡ primer, ÁÖÇü»ùÇÃ, H2O¸¸ ³Ö¾îÁÖ¸é °£´ÜÇÏ°Ô ½ÇÇèÀ» ÁøÇàÇÒ ¼ö ÀÖ´Ù.

¾Æ·¡´Â ½ÃÁß¿¡ ÆǸÅÁßÀÎ 4°³ÀÇ multiplex PCR ½Ã¾à°ú SuperPlex™ PremixÀÇ È¿À²À» ºñ±³ÇÑ µ¥ÀÌÅÍÀÌ´Ù. SuperPlex™ Premix°¡ ´Ù¸¥ Á¦Ç°°ú ºñ±³Çؼ­ ªÀº ½Ã°£ ¾È¿¡ °¡Àå ÀÏÁ¤ÇÑ È¿À²ÀÇ ÁõÆøÀ» º¸¿©Áشٴ °ÍÀ» È®ÀÎÇÒ ¼ö ÀÖ´Ù.

¡á Results
SuperPlex Premix provides more uniform multiplex PCR amplification than other premixes
½ÃÁß¿¡ ÆǸŵǴ 4°³ÀÇ ´Ù¸¥ multiplex PCR¿ë ½Ã¾à°ú SuperPlex™ Premix (Code 638543)ÀÇ È¿À²À» ºñ±³ÇÏ¿´´Ù (20-plex ¹ÝÀÀ). SuperPlex™ Premix°¡ ¸ðµç Å©±âÀÇ Å¸°Ù¿¡¼­ (human CFTR À¯ÀüÀÚÀÇ 20°³ Ÿ°Ù, 70~697 bp) °¡Àå ÀÏÁ¤ÇÑ ÁõÆø È¿À²À» º¸¿´°í, Àü±â¿µµ¿ °á°ú gel¿¡¼­ ¸ðµç ¹êµå°¡ ¼±¸íÇÏ°Ô È®ÀεǾú´Ù (±×¸² 1. Lane 1). ´Ù¸¥ ¼¼ °³ÀÇ Å¸»ç master mix (KAPA (Lane 2), Invitrogen (Lane 3)¿Í µÎ°¡Áö ¹ÝÀÀÁ¶°ÇÀÇ Qiagen (Lane 5, 6)) ¶ÇÇÑ 20°³ÀÇ Å¸°ÙÀ» ÁõÆøÇÒ ¼ö ÀÖ¾ú´Ù. ÇÏÁö¸¸, ¿©·¯ primerµéÀÇ ÁõÆø È¿À²ÀÌ ÀÏÁ¤ÇÏÁö ¾Ê°í Å©±â°¡ Å©°Å³ª ÀÛÀº Ÿ°Ù¿¡¼­ ÁõÆø È¿À²ÀÌ ³·¾ÆÁ³´Ù. ¶ÇÇÑ NEBÀÇ multiplex master mix (Lane 4)´Â 20-plexÀÇ ÁõÆøÀÌ ºÒ°¡´ÉÇÏ¿´´Ù.

µû¶ó¼­, SuperPlex™ Premix (Code 638543)À» »ç¿ëÇϸé ÀÏÁ¤ÇÑ ÁõÆø È¿À²À» Á¦°øÇÏ´Â °Í»Ó¸¸ ¾Æ´Ï¶ó ÃÖ¼ÒȯÀÇ ÃÖÀûÈ­ °úÁ¤À¸·Î ¼ö¸¹Àº primer setÀÇ multiplex PCRÀ» ÁøÇàÇÒ ¼ö ÀÖ´Ù´Â °ÍÀ» È®ÀÎÇÒ ¼ö ÀÖ¾ú´Ù.

Lane

Premix

M

Molecular weight marker (50–2,000 bp)

1

SuperPlex™ Premix

2

KAPA2G Fast Multiplex Mix

3

Invitrogen Platinum Multiplex PCR Master Mix

4

NEB Multiplex PCR 5X Master Mix

5

Qiagen Multiplex PCR Kit, without Q-solution

6

Qiagen Multiplex PCR Kit, with Q-solution

±×¸² 1. SuperPlex™ Premix¸¦ ÀÌ¿ëÇÑ 20-plex multiplex ¹ÝÀÀ¿¡¼­ÀÇ ÀÏÁ¤ÇÑ ÁõÆø È¿À²
Multiplex PCRÀº Á¦Á¶»çÀÇ Ãßõ Á¶°Ç¿¡ µû¶ó ÁøÇàµÇ¾ú´Ù. °¢ laneÀº ´Ù¸¥ PCR premix¿¡ ÀÇÇØ ÇÕ¼ºµÈ PCR »ê¹°À̸ç Àü±â¿µµ¿ ÈÄ EtBr·Î ¿°»öÇÏ¿© °á°ú¸¦ È®ÀÎÇÏ¿´´Ù.

SuperPlex Premix provides faster results than other multiplex PCR premixes
Multiplex PCRÀº °¡Àå È¿À²ÀûÀÎ ¹æ¹ýÀ¸·Î °¡Àå ¸¹Àº ¾çÀÇ µ¥ÀÌÅ͸¦ ¾ò±â À§ÇØ ÁøÇàÇÏ´Â ½ÇÇè¹ýÀ̹ǷΠSuperPlex™ Premix (Code 638543)´Â ¹ÝÀÀ setup ½Ã°£°ú Àüü ½ÇÇè ½Ã°£À» ÃÖ¼ÒÈ­Çϵµ·Ï Á¦À۵Ǿú´Ù. ¾Æ·¡ÀÇ ºñ±³ÀڷḦ º¸¸é SuperPlex™ Premix°¡ Ÿ»çÁ¦Ç° ´ëºñ PCR¿¡ °¡Àå ªÀº ½Ã°£ÀÌ ¼Ò¿äµÇ´Â °ÍÀ» È®ÀÎÇÒ ¼ö ÀÖ´Ù

 


SuperPlex Premix

KAPA2G Fast Multiplex Mix

Invitrogen Platinum Multiplex PCR Master Mix

NEB Multiplex PCR 5X Master Mix

Qiagen Multiplex PCR Kit, without Q-solution

Qiagen Multiplex PCR Kit, with Q-solution

# PCR cycles recommended

30

30

30

30

35

35

Total recommended cycle time

18ºÐ 30ÃÊ

48ºÐ

92ºÐ

71ºÐ

137ºÐ 30ÃÊ

137ºÐ 30ÃÊ

Ç¥ 1. °¢ Á¦Á¶»çÀÇ Ãßõ PCR ¹ÝÀÀ ½Ã°£
SuperPlex™ Premix´Â 20-plex¿¡¼­ °¡Àå ªÀº ¹ÝÀÀ½Ã°£À» °¡Áø multiplex PCR È¿¼ÒÀÌ´Ù.

°¢ multiplex PCR¿ë ½Ã¾àÀ¸·Î human CFTR À¯ÀüÀÚÀÇ 20°³ Ÿ°ÙÀ» PCR cycles ½Ã°£ÀÌ °¡Àå ªÀº SuperPlex™ Premix (Code 638543)ÀÇ ¹ÝÀÀÁ¶°Ç¿¡¼­ Å×½ºÆ®ÇÏ¿´´Ù (±×¸² 2.). ±× °á°ú, SuperPlex™ Premix°¡ °¡Àå ¶Ù¾î³­ È°¼ºÀ» º¸¿´À¸¸ç, NEB¿Í Qiagen master mixÀÇ °æ¿ì ÁõÆø¿¡ ½ÇÆÐÇÏ¿´°í (Lane 4~6), KAPA2G ¿Í InvitrogenÀÇ master mix´Â ³·Àº È°¼º°ú smearing ¹× amplicon dropoutÀÌ È®ÀεǾú´Ù (Lane 2~3).

Lane

Premix

M

Molecular weight marker (50–2,000 bp)

1

SuperPlex™ Premix

2

KAPA2G Fast Multiplex Mix

3

Invitrogen Platinum Multiplex PCR Master Mix

4

NEB Multiplex PCR 5X Master Mix

5

Qiagen Multiplex PCR Kit, without Q-solution

6

Qiagen Multiplex PCR Kit, with Q-solution

±×¸² 2. ªÀº ¹ÝÀÀ½Ã°£¿¡µµ SuperPlex™ PremixÀÇ ¼º°øÀûÀÎ ÁõÆø (20-plex multiplex)
Ç¥ 4¿¡ ¸í½ÃµÈ SuperPlex™ PremixÀÇ ÂªÀº PCR cycle Á¶°Ç¿¡¼­ SuperPlex™ Premix¿Í Ÿ»ç multiplex PCR premix ½Ã¾àÀÇ È¿À²À» ºñ±³Çß´Ù. °¢ laneÀº ´Ù¸¥ PCR premix¿¡ ÀÇÇØ Á¦ÀÛµÈ PCR »ê¹°À̸ç Àü±â¿µµ¿ ÈÄ °á°ú´Â EtBrÀ» ÅëÇØ ¿°»öµÇ¾ú´Ù.

¡á Conclusions
½ÇÇè °á°ú, SuperPlex™ Premix (Code 638543)´Â ½ÃÁß¿¡ ´Ù¸¥ multiplex PCR premix ½Ã¾à°ú ºñ±³Çؼ­ ÀÏÁ¤ÇÑ È¿À²°ú ºü¸¥ ÁõÆø¼Óµµ¸¦ °¡Áø ½Ã¾àÀ¸·Î, CFTR À¯ÀüÀÚ¸¦ Ÿ°ÙÇÏ´Â 20-plexÀÇ PCR »ê¹°ÀÌ ¸ðµÎ ÀÏÁ¤ÇÑ ÁõÆø È¿À²À» º¸ÀÌ¸ç °¡Àå ªÀº ½ÇÇè½Ã°£ÀÌ ¼Ò¿äµÇ¾ú´Ù.

µû¶ó¼­, SuperPlex™ Premix´Â ¼Óµµ, ÁõÆøÀÇ ÀÏ°ü¼º, »ç¿ë ÆíÀǼº¿¡¼­ °­Á¡ÀÌ ÀÖ´Â ÃÖÀûÀÇ multiplex PCR ½Ã¾àÀ̶ó°í ÇÒ ¼ö ÀÖ´Ù.

¡á Method
Multiplex PCR¿¡¼­ SuperPlex™ Premix (Code 638543)ÀÇ È¿À²À» È®ÀÎÇϱâ À§ÇØ CFTR À¯ÀüÀÚ¿¡¼­ ¾Æ·¡¿Í °°Àº 20°³ setÀÇ primer¸¦ »ç¿ëÇÏ¿© multiplex PCRÀ» ÁøÇàÇÏ¿´´Ù. Primer´Â ÃÖÁ¾ PCR »ê¹°ÀÇ Å©±â°¡ 70~697 bp°¡ µÇµµ·Ï µðÀÚÀεǾú´Ù. ÀÌÁß 15 ¼¼Æ®ÀÇ primer´Â ÀÌÀü ¿¬±¸ (Shuber et al. 1995)¿¡¼­ »ç¿ëµÇ¾ú´ø primerÀÌ°í 5 ¼¼Æ®´Â Primer3 tool·Î »õ·Ó°Ô µðÀÚÀÎÇß´Ù. PCR ¹ÝÀÀÀº »ç¶÷ÀÇ gDNA 10 ngÀ» ÁÖÇüÀ¸·Î »ç¿ëÇÏ°í, ÀÌÈÄ PCR »ê¹° 18 §¡¸¦ 1x TAE ¹öÆÛ·Î Á¦ÀÛµÈ 4% agarose gel¿¡¼­ 80 V·Î 200ºÐ°£ Àü±â¿µµ¿ÇÑ ÈÄ EtBr·Î ¿°»öÇÏ¿© È®ÀÎÇÏ¿´´Ù.

Target

Forward primer sequence

Reverse primer sequence

Reference

CFTR intron 19 [22]

GAA TCA TTC AGT GGG TAT AAG CAG

AGG CTT CTC AGT GAT CTG TTG

Shuber et al. 1995

CFTR exon 19 [22]

AGT CTA ACA AAG CAA GCA GTG

GCC CGA CAA ATA ACC AAG TGA

Shuber et al. 1995

CFTR exon 21 [24]

CAA AAG TAC CTG TTG CTC CA

TGA TGG TAA GTA CAT GGG TG

Shuber et al. 1995

CFTR exon 9 [10]

CCA CTG AAA ATA ATA TGA GGA AAT

CTT CTA ATG GTG ATG ACA GCC T

Shuber et al. 1995

CFTR exon 13 [14]

TAA GGG AGT CTT TTG CAC AA

AGG TAG CAG CTA TTT TTA TGG

Shuber et al. 1995

CFTR exon 4

CGA TAC AGA ATA TAT GTG CC

TGT AGG AAG TCA CCA AAG

Shuber et al. 1995

CFTR exon 17b [20]

AGT TAA TGA GTT CAT AGT ACC TGT T

GGA GTC CAA TTT TCA CTC ATC TTG

Shuber et al. 1995

CFTR exon 7 [8]

GGT ACA TTA CCT GTA TTT TGT TT

AGA TAC TTC AAT AGC TCA GCC

Shuber et al. 1995

CFTR exon 11 [12]

TAC ATG AAT GAC ATT TAC AGC A

CAG ATT GAG CAT ACT AAA AGT G

Shuber et al. 1995

CFTR exon 10 [11]

TCA CAT AGT TTC TTA CCT CT

GAG CCT TCA GAG GGT AAA AT

Shuber et al. 1995

CFTR exon 20 [23]

TCC TTT TGC TCA CCT GTG GT

AAG AAC TGG ATC AGG GAA GA

Shuber et al. 1995

CFTR exon 5

GTA TAA TTT ATA ACA ATA GTG CC

GCT GTC AAG CCG TGT TCT A

Shuber et al. 1995

CFTR exon 14b [16]

ACA ATA CAT ACA AAC ATA GTG G

TTG GTT GTG CTG TGG CTC CT

Shuber et al. 1995

CFTR exon 12 [13]

GCA TGA GCA TTA TAA GTA AGG

GAC TCT CCT TTT GGA TAC CTA

Shuber et al. 1995

CFTR exon 3

ACA AAT GAG ATC CTT ACC CC

GGC GAT GTT TTT TCT GGA GA

Shuber et al. 1995

CFTR intron 10b

CTG TCA AGG CTG TTC TCA CT

TTG CCC ATC CTA TTC TCG TG

½Å±Ô µðÀÚÀÎµÈ primer

CFTR intron 6b - 526

GAT CAT CAC ACC AGA GCC TTA G

CAC TGA TCT TCC CAG CTC TC

½Å±Ô µðÀÚÀÎµÈ primer

CFTR intron 6a - 572

CCA GGA CTG CTC TAT GCA TAG

GAT CCA TAA ACC AGA GTA ATG AAA GCT

½Å±Ô µðÀÚÀÎµÈ primer

CFTR intron 7a - 638

CTG TCA ATA TTC ATG CAT TAG TTG CAC

GAT TGA AGT TTG TAG AAV ATG GAG CTC

½Å±Ô µðÀÚÀÎµÈ primer

CFTR intron 3c

AGG GAT AGG AGA ATG GAG TAT GTA T

GTG TTA CAG TAG TTT CAA GAA TCC C

½Å±Ô µðÀÚÀÎµÈ primer


Ç¥ 2. Multiplex PCR¿¡¼­ »ç¿ëµÈ primer
À§ÀÇ primer¿¡´Â UPS ÅÂ±× ¼­¿­·Î GGTTCAGT ¼­¿­ÀÌ ¸ðµç PrimerÀÇ 5’-end¿¡ Ãß°¡µÇ¾î ÀÖ´Ù (À§ÀÇ ¼­¿­¿¡´Â Æ÷½ÃµÇ¾î ÀÖÁö ¾ÊÀ½)

Experiment 1: testing PCR amplification uniformity
ù ½ÇÇèÀº °¢ Á¦Á¶»ç¿¡¼­ ¾È³»ÇÏ´Â ½ÇÇè Á¶°Ç (Ç¥ 3)À¸·Î 20-plex PCRÀÇ ÁõÆøÈ¿À²À» Å×½ºÆ®ÇÏ¿´´Ù. SuperPlex Premix´Â PCR »ê¹°ÀÇ Å©±â°¡ 300 bp ÀÌÇÏÀÎ °æ¿ì primer´Â ÃÖÁ¾ ³óµµ°¡ 300 nMÀÌ µÇµµ·Ï »ç¿ëµÇ¾ú°í, PCR »ê¹°ÀÇ Å©±â°¡ 300 bp ÀÌ»óÀÎ °æ¿ì primer´Â ÃÖÁ¾ ³óµµ°¡ 150 nMÀÌ µÇµµ·Ï »ç¿ëÇÏ¿´´Ù. ¶ÇÇÑ Å¸»çÀÇ master mix Á¦Ç°Àº °¢ Á¦Á¶»ç¿¡¼­ ¾È³»ÇÏ´Â primer ³óµµ¸¦ »ç¿ëÇÏ¿© ½ÇÇèÇÏ¿´´Ù (KAPA2G, NEB, Qiagen master mixes - 200 nM; Invitrogen 100 nM).

Lane

 Premix

Step

¿Âµµ

SuperPlex™ Premix

KAPA2G Fast Multiplex Mix

Invitrogen Platinum Multiplex PCR Master Mix

NEB Multiplex PCR 5X Master Mix

Qiagen Multiplex PCR Kit

Initial

denaturation

95¡É

2 min

3 min

2 min

1 min

15 min

Denaturation

95¡É

10 sec

15 sec

30 sec

20 sec

30 sec (94¡É)

Annealing

60¡É

10 sec

30 sec

1 min 30 sec

1 min

1 min 30 sec

Extension

72¡É

15 sec

15 sec

1 min

1 min

1 min 30 sec

# PCR cycles

 

30

30

30

30

35

Final extension

72¡É

N/A

1 min

10 min

5 min (68¡É)

10 min

Ç¥ 3. 20-plex PCR ¹ÝÀÀ¿¡ 1Â÷Ãßõ PCR Á¶°Ç
½ÇÇèÀº °¢ Á¦Á¶»ç¿¡¼­ ±ÇÀåÇÏ´Â ½ÇÇè Á¶°Ç¿¡ µû¶ó Å×½ºÆ®µÇ¾ú´Ù. ¶ÇÇÑ Qiagen Multiplex PCR Kit´Â °°Àº Á¶°Ç¿¡¼­ Q-solution ÷°¡ ¿©ºÎ¿¡ µû¶ó µÎ°¡Áö ¹æ¹ýÀ¸·Î Å×½ºÆ®µÇ¾ú´Ù.

Experiment 2: testing PCR cycle times
µÎ ¹ø° ½ÇÇèÀº ½ÃÁß¿¡ ÆǸÅÁßÀÎ multiplex PCR¿ë ½Ã¾àÀ» »ç¿ëÇØ SuperPlex¢â PremixÀÇ PCR cycle·Î (Ç¥ 4) ´õ ªÀº ¹ÝÀÀ½Ã°£¿¡¼­ÀÇ 20-plex PCR È¿À²À» È®ÀÎÇÏ¿´´Ù. ¸ðµç ½Ã¾à¿¡¼­ primer ³óµµ´Â PCR »ê¹°ÀÇ Å©±â°¡ 300 bp ÀÌÇÏÀ϶§ ÃÖÁ¾ ³óµµ 300 nMÀ¸·Î, PCR »ê¹°ÀÇ Å©±â°¡ 300 bp ÀÌ»óÀÏ ¶§ ÃÖÁ¾ ³óµµ 150 nMÀÌ µÇµµ·Ï »ç¿ëÇÏ¿´´Ù.

Ç¥ 4. ½ÇÇè 2¿¡¼­ÀÇ SuperPlex™ Premix¿Í ½ÃÁßÀÇ multiplex PCR¿ë ½Ã¾àÀ» Å×½ºÆ®ÇÑ PCR Á¶°Ç

Reference
Shuber A. P., Grondin V. J. & Klinger K. W. A simplified procedure for developing multiplex PCRs. Genome Res5, 488–493 (1995).