EasyClone cDNA library construction kit

Kit for construction of directional cDNA libraries from total or polyA+ RNA

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Article number: P01010 Size: 20 reactions Storage temperature: -20°C Shipping temperature: -20°C

Applications

Directional cDNA library construction
Compatible with DUALhybrid (yeast two-hybrid), DUALmembrane and DUALhunter systems

Benefits

Fast and easy synthesis of highly complex, representational cDNAs
High representation of full length transcripts
Directional cloning of cDNAs into DUALhybrid (yeast two-hybrid), DUALmembrane and DUALhunter vectors
Only small starting amounts of RNA required
Sufficient reagents to construct 10 libraries

EasyClone cDNA synthesis is based on a novel technology utilizing the specific features of MMLV-based reverse transcriptase to create full length enriched double stranded cDNA from total or polyA+ RNA samples. cDNAs are amplified by representational PCR and cloned directionally into DUALhybrid, DUALmembrane or DUAlhunter library vectors.

No extensive know-how in library construction is needed - the kit supplies all reagents needed for library construction and the detailed manual will guide you through all steps of construction.

cDNA library construciton service procedure

Box 1

5X first strand buffer	80 µl
DTT (20 mM)	30 µl
10X dNTP mix (10 mM each)	25 µl
PlugOligo-1 adapter (15 µM)	25 µl
CDS-1 adapter (10 µM)	25 µl
PlugOligo-3M adapter (15 µM)	25 µl
CDS-3M adapter (10 µM)	25 µl
EasyClone reverse transcriptase	20 µl
IP-solution	130 µl
Control RNA template (0.5 µg/µl)	15 µl
Sterile RNase free water	1.8 ml

Box 2

50X EasyClone polymerase mix :	50 µl
10X EasyClone buffer	300 µl
PCR Primer M1 (10 µM)	100 µl
50X dNTP mix (10 mM each)	80 µl
Sterile RNase free water	1.8 ml
Control amplified cDNA sample 1	25 µl
Control amplified cDNA sample 2	25 µl