n-Fusion 2019-2020 In-Fusion P.3
In-Fusion P.4
P.15, 16
P.17, 18
P.19

In-Fusion50 15
PCR

OK
In-Fusion®
DNA
kit
N T M K


3
In-Fusion - 2 -
Insert Cloning Protocol-At-A-Glance

E. coli HST08 Premium Competent Cells 9128 Stellar™ Competent Cells 636763 DNA10 kb
PCR 155’
In-Fusion

10 kb 10 kb 0.5 kb 0.510 kb 10 kb 50100 ng 50200 ng 1050 ng 50100 ng 50200 ng
1 2PCR
- 3 - In-Fusion
DNA
In-Fusion15
In-FusionPCR
In-Fusion
In-Fusion

PrimeSTAR® GXL
250 U R050A ¥37,000
PrimeSTAR® Max
100 R045A ¥31,500
PrimeSTAR®
50 μl
CRISPRIn-Fusion® HD Cloning KitCas9 CRISPR-mediated modular RNA-guided regulation of transcription in eukaryotes. Gilbert LA et al., Cell, (2013) Jul 18; 154(2):442-451
4In-Fusion® HD Cloning Kit High-throughput tetrad analysis. Ludlow CL. et al., Nat Methods. (2013) Jul; 10(7):671-675

In-Fusion
1 1kb
In-Fusion® HD Cloning Kit4010 PCR3 B1PCR3

In-Fusion® HD Cloning Kit
1 2 3 4 5
M a b c a b c a b c a b c a b c
40
1
M:λ-Hind III digest
PCR

pUC19 DNA 3219BamH IIn-Fusion® HD Cloning Kit1,530 bp726 bp 613 bp2,906 bp4
1005PCR2

8,461 bp
Marker 1 kb ladder
1
2
M 1 2 3 4 5 6 7 8 9 10
M 1 2 3 4 5 6 7 8 9 10
X-Gal/IPTG10 12 9/107/10DNA
4
In-Fusion® Cloning Kit
- 9 - In-Fusion
169 56 60

Cloning EnhancerIn-Fusion In-Fusion
PCR In-Fusion101

In-Fusion - 10 -
6
In-Fusion® Cloning Kit4
4PCR43
T5p 169 bp blc-bamM 2,199 bp rrnBT1terminator 131 bp lacIp-lacI 1,161 bp
In-Fusion
In-Fusion
- 11 - In-Fusion
7
75 bpSfi I, Pml I5,900 bp 50 ng 020 ng10 μl 1/101/10 In-Fusion® HDAB

vector 50 ng insertng
0 0.5 1 2 5 10 20

In-Fusion
In-Fusion
In-Fusion - 12 -
SfiI PmlI
Sfi I Pml I
70010EcoR I1% 750 bp
In-Fusion
In-Fusion
- 13 - In-Fusion
In-Fusion® HD
PCR product
750 bp
Linearized vector
4,700 bp
9
P2A
72
Marker 1 kb ladder
PCRcloning enhancerIn- Fusion100%

site-directed mutagenesis PCR
100%In-Fusion In-Fusion1
H T T K
T Y N K
PCR product In-Fusion
K I S A
K A S Y
1 insertvector1
5ligation
ligase 1
1
T I T Y
N T K K
1
2.7 kbMBP1.1 kbPROF120.7 kbAcGFP0.7 kb3DNA
In-Fusion® HD Cloning A
Vector + Insert 89 111 392
Negative Control no insert 1 39 78
10026/26 195 /26 7319 /26
2.7 kb1MBP 1.1 kb
In-Fusion® HD Cloning A
Vector + Insert 635 401

2





PrimeSTAR®PCR
96In-Fusion®11A 711 In-Fusion®
In-Fusion®A


pUC19PrimeSTAR® GXL DNA Polymerase R050A/BPCR PrimeSTAR® Max DNA Polymerase R045/BPCR 15 bp 5015 Stellar™ Competent Cells 636763 96



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100 639635 ¥210,000
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In-Fusion® HD Cloning
In-Fusion
In-Fusion® HD EcoDry™ Cloning Kit In-Fusion® HD
Cloning Enhancer
NucleoSpin® Gel and PCR Clean-up
PCR
CloneAmp™ HiFi PCR Premix
In-Fusion® HD
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