AccuPower® qPCR Array System: Single gene qPCR primer (200 reactions)

AccuPower® qPCR Array System: Single gene qPCR primer는 qPCR array system의 qPCR primer를 200 rxn씩 제공해 드리는 제품입니다. Single gene qPCR primer는 target specificity 및 PCR efficiency가 검증되어 별도의 검증 없이 실험에 바로 이용하실 수 있습니다.

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카탈로그 번호
S-6042-S200

Reference gene(internal control gene)은 주로 세포 생존에 필수적인 유전자들이며, 모든 조직에서 일정하게 발현된다고 알려져 있습니다. 하지만 실험 조건, 세포 종 등에 의한 차이로 발현량이 변하기도 합니다. qPCR을 이용한 상대정량 분석법 (principles of relative quantification)은 reference group과 목표 유전자의 발현량을 상대적으로 비교하는 방법입니다. 상대정량 분석법인 2-ddCt method는 reference gene을 사용하여 extraction yield, reverse transcription yield, amplification efficiency 그리고 사용된 RNA의 량을 normalize해주고 있습니다. 선별된 reference gene은 반드시 안정적으로 발현되어야 하고 sample의 total RNA량과 밀접하게 연관되어야 합니다.

  

Figure 1. ACTB expression levels in the various tissues. (biogps.org)

AccuPower® qPCR Array System: Human Reference qPCR primer A set
# Gene symbol Accession # Description Also known as
A ACTB NM_001101 Actin, beta BRWS1, PS1TP5BP1
B B2M NM_004048 Beta-2-microglobulin -
C GAPDH NM_001256799
NM_001289745
NM_001289746
NM_002046
Glyceraldehyde-3-phosphate dehydrogenase G3PD, GAPD, HEL-S-162eP
D GUSB NM_000181
NM_001284290
NM_001293104
NM_001293105
Glucuronidase, beta BG, MPS7
E HPRT1 NM_000194 Hypoxanthine phosphoribosyltransferase 1 HPRT, HGPRT
F PPIA NM_001300981
NM_021130
Peptidylprolyl isomerase A (cyclophilin A) CYPA, CYPH, HEL-S-69p
G RPL13A NM_001270491
NM_012423
Ribosomal protein L13a L13A, TSTA1
H RPLP0 NM_001002
NM_053275
Ribosomal protein, large, P0 P0, LP0, L10E, RPP0, PRLP0

AccuPower® qPCR Array System: Human Reference qPCR primer B set
# Gene symbol Accession # Description Also known as
I ALAS1 NM_000688
NM_001304443
NM_001304444
NM_199166
5'-aminolevulinate synthase 1 ALAS, MIG4, ALAS3, ALASH, ALAS-H
J H6PD NM_001282587
NM_004285
Hexose-6-phosphate dehydrogenase
(glucose 1-dehydrogenase)
GDH, G6PDH, CORTRD1
K PGK1 NM_000291 Phosphoglycerate kinase 1 PGKA, MIG10, HEL-S-68p
L TBP NM_001172085
NM_003194
TATA box binding protein HDL4, GTF2D, SCA17, TFIID, GTF2D1
M TFRC NM_001128148
NM_003234
Transferrin receptor T9, TR, TFR, p90, CD71, TFR1, TRFR
N TUBB NM_001293212
NM_001293213
NM_001293214
NM_001293215
NM_001293216
NM_178014
Tubulin, beta class I M40, TUBB1, TUBB5, CDCBM6, OK/SW-cl.56
O UBC NM_021009 Ubiquitin C HMG20
P YWHAZ NM_001135699
NM_001135700
NM_001135701
NM_001135702
NM_003406
NM_145690
Tyrosine 3-monooxygenase/tryptophan
5-monooxygenase activation protein, zeta
HEL4, YWHAD, KCIP-1, HEL-S-3, 14-3-3-zeta

AccuPower® qPCR Array System: Mouse Reference qPCR primer A set
# Gene symbol Accession # Description Also known as
a Actb NM_007393 Actin, beta Actx, beta-actin, E430023M04Rik
b B2m NM_009735 Beta-2 microglobulin Ly-m11, beta2m, beta2-m
c Gapdh NM_001289726
NM_008084
Glyceraldehyde-3-phosphate dehydrogenase Gapd
d Gusb NM_010368 Glucuronidase, beta Gur, Gus, Gut, asd, Gus-r, Gus-s, Gus-t, Gus-u, AI747421
e Hprt NM_013556 Hypoxanthine guanine phosphoribosyl transferase HPGRT, Hprt1, C81579
f Ppia NM_008907 Peptidylprolyl isomerase A Cphn, CypA, CyP-18, 2700098C05
g Rpl13a NM_009438 Ribosomal protein L13A Tstap198-7, tum-antigen, 1810026N22Rik
h Tbp NM_013684 TATA box binding protein Gtf2d, SCA17, TFIID, GTF2D1

AccuPower® qPCR Array System: Mouse Reference qPCR primer A set
# Gene symbol Accession # Description Also known as
i Canx NM_001110499
NM_001110500
NM_007597
Calnexin Cnx, AI988026, D11Ertd153e, 1110069N15Rik
j Cyc1 NM_025567 Cytochrome c-1 AA408921, 2610002H19Rik
k Hsp90ab1 NM_008302 Heat shock protein 90 alpha (cytosolic),
class B member 1
90kDa, Hsp84, Hsp90, Hspcb, C81438, Hsp84-1, AL022974
l Ldhal6b NM_175349 Lactate dehydrogenase A-like 6B LDHL, Ldhal, AI326310, 4933402O15Rik
m Sdha NM_023281 Succinate dehydrogenase complex, subunit A, flavoprotein (Fp) FP, SDH2, SDHF, C81073, 4921513A11, 1500032O14Rik, 2310034D06Rik
n Tfrc NM_011638 Transferrin receptor TR, TFR, p90, CD71, TFR1, Trfr, Mtvr1, Mtvr-1, AI195355, AI426448, AU015758, 2610028K12Rik, E430033M20Rik
o Ubc NM_019639 Ubiquitin C Ubb, Uba52, Rps27a, TI-225, AI194771, 2700054O04Rik
p Ywhaz NM_001253805
NM_001253806
NM_001253807
NM_011740
Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, zeta HEL4, YWHAD, KCIP-1, HEL-S-3, 14-3-3-zeta

Applications

Applicable to Quantitative Real-time PCR


Primer specificity

• Primer blast 및 자사 bioinformatics tool을 이용한 타겟 특이적인 primer 디자인
• Self primer-dimer 형성 서열 배제
• Dissociation curve에서 단일 peak 확인
• Gel 전기영동에서 단일밴드 확인
• 70~150 bp의 짧은 amplicon size



Figure 1. Primer validation result (Example. RPL13A gene)

Wide linear dynamic range & high PCR efficiency

• 102 ~ 107 copies의 광범위한 증폭 범위 • MIQE Guide line을 준수하는 90 ~ 110%의 qPCR 증폭 효율

1) PCR efficiency test
   : Purified qPCR product를 10배씩 serial dilution 하여 efficiency test.


Figure 2. [BIONEER] Dynamic range & PCR efficiency (Example. GAPDH gene).
MIQE guideline에서 권고하는 qPCR efficiency 범위(100%±10%)에 해당.
Limit of detection (LOD) 100 copies 이하 확인.


Figure 3. [Competitor] Dynamic range & PCR efficiency (Example. GAPDH gene).
MIQE guideline에서 권고하는 qPCR efficiency 범위(100%±10%)에 해당.
103과 102 copies 사이에 불규칙적인 간격이 관찰되며 이는 limit of detection (LOD) 값이
103 copies 이하로 자사 대비 detection 효율이 떨어짐.


2) Target specificity test
   : Purified qPCR product를 10배씩 serial dilution + reference mouse cDNA (1㎍ mouse total RNA/20㎕)로 efficiency test.

Figure 4. [BIONEER] Mouse template를 이용한 target specificity test (Example. ACTB gene).
표적 유전자에 대한 specificity와 MIQE guideline에서 권고하는 qPCR efficiency 범위 (100%±10%)에 해당.
Limit of detection (LOD) 100 copy 이하 확인.


Figure 5. [Competitor] Mouse template를 이용한 target specificity test (Example. ACTB gene).
104 copy부터 efficiency fluorescence curve의 뭉침 현상 확인.
MIQE guideline에서 권고하는 qPCR efficiency 범위(100%±10%)에 벗어난 161% efficiency를 보이며, 자사 대비 target specificity가 떨어짐을 확인.

  

Gene List

AccuPower® qPCR Array System: Human Reference qPCR primer A set
# Gene symbol Accession # Description Also known as
A ACTB NM_001101 Actin, beta BRWS1, PS1TP5BP1
B B2M NM_004048 Beta-2-microglobulin -
C GAPDH NM_001256799
NM_001289745
NM_001289746
NM_002046
Glyceraldehyde-3-phosphate
dehydrogenase
G3PD, GAPD, HEL-S-162eP
D GUSB NM_000181
NM_001284290
NM_001293104
NM_001293105
Glucuronidase, beta BG, MPS7
E HPRT1 NM_000194 Hypoxanthine phosphoribosyltransferase 1 HPRT, HGPRT
F PPIA NM_001300981
NM_021130
Peptidylprolyl isomerase A (cyclophilin A) CYPA, CYPH, HEL-S-69p
G RPL13A NM_001270491
NM_012423
Ribosomal protein L13a L13A, TSTA1
H RPLP0 NM_001002
NM_053275
Ribosomal protein, large, P0 P0, LP0, L10E, RPP0, PRLP0

AccuPower® qPCR Array System: Human Reference qPCR primer B set
# Gene symbol Accession # Description Also known as
I ALAS1 NM_000688
NM_001304443
NM_001304444
NM_199166
5'-aminolevulinate synthase 1 ALAS, MIG4, ALAS3, ALASH, ALAS-H
J H6PD NM_001282587
NM_004285
Hexose-6-phosphate dehydrogenase
(glucose 1-dehydrogenase)
GDH, G6PDH, CORTRD1
K PGK1 NM_000291 Phosphoglycerate kinase 1 PGKA, MIG10, HEL-S-68p
L TBP NM_001172085
NM_003194
TATA box binding protein HDL4, GTF2D, SCA17, TFIID, GTF2D1
M TFRC NM_001128148
NM_003234
Transferrin receptor T9, TR, TFR, p90, CD71, TFR1, TRFR
N TUBB NM_001293212
NM_001293213
NM_001293214
NM_001293215
NM_001293216
NM_178014
Tubulin, beta class I M40, TUBB1, TUBB5, CDCBM6, OK/SW-cl.56
O UBC NM_021009 Ubiquitin C HMG20
P YWHAZ NM_001135699
NM_001135700
NM_001135701
NM_001135702
NM_003406
NM_145690
Tyrosine 3-monooxygenase/tryptophan
5-monooxygenase activation protein, zeta
HEL4, YWHAD, KCIP-1, HEL-S-3, 14-3-3-zeta

AccuPower® qPCR Array System: Mouse Reference qPCR primer A set
# Gene symbol Accession # Description Also known as
a Actb NM_007393 Actin, beta Actx, beta-actin, E430023M04Rik
b B2m NM_009735 Beta-2 microglobulin Ly-m11, beta2m, beta2-m
c Gapdh NM_001289726
NM_008084
Glyceraldehyde-3-phosphate
dehydrogenase
Gapd
d Gusb NM_010368 Glucuronidase, beta Gur, Gus, Gut, asd, Gus-r, Gus-s, Gus-t, Gus-u, AI747421
e Hprt NM_013556 Hypoxanthine guanine phosphoribosyl transferase HPGRT, Hprt1, C81579
f Ppia NM_008907 Peptidylprolyl isomerase A Cphn, CypA, CyP-18, 2700098C05
g Rpl13a NM_009438 Ribosomal protein L13A Tstap198-7, tum-antigen, 1810026N22Rik
h Tbp NM_013684 TATA box binding protein Gtf2d, SCA17, TFIID, GTF2D1

AccuPower® qPCR Array System: Mouse Reference qPCR primer A set
# Gene symbol Accession # Description Also known as
i Canx NM_001110499
NM_001110500
NM_007597
Calnexin Cnx, AI988026, D11Ertd153e, 1110069N15Rik
j Cyc1 NM_025567 Cytochrome c-1 AA408921, 2610002H19Rik
k Hsp90ab1 NM_008302 Heat shock protein 90 alpha (cytosolic),
class B member 1
90kDa, Hsp84, Hsp90, Hspcb, C81438, Hsp84-1, AL022974
l Ldhal6b NM_175349 Lactate dehydrogenase A-like 6B LDHL, Ldhal, AI326310, 4933402O15Rik
m Sdha NM_023281 Succinate dehydrogenase complex,
subunit A, flavoprotein (Fp)
FP, SDH2, SDHF, C81073, 4921513A11, 1500032O14Rik, 2310034D06Rik
n Tfrc NM_011638 Transferrin receptor TR, TFR, p90, CD71, TFR1, Trfr, Mtvr1, Mtvr-1, AI195355, AI426448, AU015758, 2610028K12Rik, E430033M20Rik
o Ubc NM_019639 Ubiquitin C Ubb, Uba52, Rps27a, TI-225, AI194771, 2700054O04Rik
p Ywhaz NM_001253805
NM_001253806
NM_001253807
NM_011740
Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, zeta HEL4, YWHAD, KCIP-1, HEL-S-3, 14-3-3-zeta

Cat. No. Product Description Price
S-6042-S200 AccuPower® qPCR Array System: Single gene qPCR primer, 200 rxns/tube ₩150,000
S-6042-TH0A AccuPower® qPCR Array System: Human Reference qPCR primer A set, 20 rxn ₩70,000
S-6042-TH0B AccuPower® qPCR Array System: Human Reference qPCR primer B set, 20 rxn ₩70,000
S-6042-TM0A AccuPower® qPCR Array System: Mouse Reference qPCR primer A set, 20 rxn ₩70,000
S-6042-TM0B AccuPower® qPCR Array System: Mouse Reference qPCR primer B set, 20 rxn ₩70,000

* VAT 별도입니다.

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