Strategi Kloning

CLONING STRATEGY

• Strategy depends on the starting information and desired

endpoint.
• Starting Information or Resources:
▪ Protein sequence
▪ Positional cloning information
▪ mRNA species / sequence
▪ cDNA libraries
▪ DNA sequence known or unknown
▪ genomic DNA libraries
▪ PCR product
 TINJAUAN UMUM

Sumber DNA
belum
diketahui diketahui

Ekspresi gen Identifikasi & Isolasi Gen

Lokasi Karakterisasi Gen

Aplikasi kloning Kloning Gen

Produksi protein pada Vektor
dan lain-lain

Transfer Gen
Ke Sel Inang/Target

Stable Penapisan Organisme Transient

Studi Lanjut Transgenik Expression Buang ?
Expression
Primrose, Twyman, and Old. 2006 Principles of Gene Manipulation. 6th Ed
 Cloning Techniques

Your
Source
PCR Library
ORF Gene
collection synthesis

Gene
Gene Gene
Golden Gate Using Restriction
Cloning Enzymes
MCS
Gene Vector Gene
Gibson TA/TOPO Cloning
Assembly

Gene Gene
Bi-Multi-cistronic Gateway
cloning cloning

Gene Gene
Infusion Ligation Independent
cloning cloning
Restriction Enzyme Cloning
Gateway Cloning
TOPO® Cloning
Gibson Assembly (Isothermal Assembly
Reaction)
 Type IIS Assembly
(Golden Gate & MoClo)
Ligation Independent Cloning
BI- or MULTI-CISTRONIC CLONING

Shaimardanova, A.A. et al. 2019