GeneCopoeia AORT-0020, AORT-0050 User Guide
All‐in‐One
Forreliablefirst‐strandcDNAsynthesisfromallRNAsources
Cat.No.AORT‐0020(20synthesisreactions)
Cat.No.AORT‐0050(50synthesisreactions)
UserManualI
™
First‐StrandcDNASynthesisKit
GeneCopoeia,Inc.
9620MedicalCenterDrive,#101
Rockville,MD20850
USA
301‐762‐0888
866‐360‐9531
inquiry@genecopoeia.com
www.genecopoeia.com
© 2009 GeneCopoeia, Inc.
All-in-One™ First-Strand cDNA Synthesis Kit
USER MANUAL I
™
All-in-One
I. Description
II. Related Products
III. Contents and Storage
IV. Preparation
V. Procedure
VI. Example
VII. Trouble Shooting Guide
VIII. Limited Use License and Warranty
I. Description
The All-in-One™ First-Strand cDNA Synthesis Kit includes a reverse transcriptase and a specialized set of
reagents designed to yield first-strand cDNA that is optimal for gene cloning, cDNA library creation and
quantitative PCR amplification. A robust experimental design delivers a universal kit that is suitable for firststrand cDNA synthesis from almost any source of RNA.
The kit uses Moloney Murine Leukemia Virus Reverse Transcriptase, RNase H Minus (M-MLV RTase (H–))
which is an RNA-dependent DNA polymerase used in cDNA synthesis with long RNA templates (>13kb).
The lack of RNase H activity is important in this application in that RNase H activity will start to degrade
template during long incubation times required for producing long cDNAs. RNase H minus RT enables
preparation of long cDNAs and libraries containing a high percentage of full-length cDNA.
II. Related Products
First-Strand cDNA Synthesis Kit
GeneCopoeia Description
All-in-One™ qPCR Mix
All-in-One™ qPCR Primers
All-in-One™ qPCR Primer Array
All-in-One™ miRNA qRT-PCR
Detection Kits
All-in-One™ miRNA qPCR Primers
OmicsLink™ Expression-Ready
ORF cDNA Clones
Endofectin™ Transfection Reagents
SYBR Green-based real-time quantitative
PCR Mix
Human, mouse and rat
primers
User specified, ready-touse primer arrays in 96well-plate format
®
Green-based Accurately quantify miRNA expression
SYBR
Human, mouse and rat
primers
20,000 human
15,000 mouse
Optimized for specific cell
types
Validated, gene-specific primers ensure
specificity and sensitivity
Reliable tools ideal for analyzing the
expression of a focused panel of genes
such as pathways, diseases or customized
gene panels
Validated for robust, reproducible and
reliable quantitation of miRNA activity
Perform a variety of applications with
expression-ready clones
Transfect efficiently and with low toxicity
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All-in-One™ First-Strand cDNA Synthesis Kit
III. Contents and Storage
Contents and storage recommendations for the All-in-One First-Strand cDNA Synthesis Kit
Cat. Nos. AORT-0020 and AORT-0050
Contents Quantity Storage temperature/ conditions
200 U/µl M-MLV Reverse
Transcriptase (RNase H-)
5X RT Reaction Buffer
25 U/µl RNase Inhibitor
are provided in the following table.
–20°C (Stable for at least 12 months)
1 x 20 µl
1 x 50 µl
1 x 100 µl
1 x 250 µl
1 x 20 µl
1 x 50 µl
Alternatively, the solution can also be
stored at -80°C in aliquots. Avoid
repeated freezing/ thawing.
–20°C (Stable for at least 12 months)
Alternatively, the solution can also be
stored at -80°C in aliquots. Avoid
repeated freezing/ thawing.
–20°C (Stable for at least 12 months)
Alternatively, the solution can also be
stored at -80°C in aliquots. Avoid
repeated freezing/ thawing.
25 mM dNTP
60 µM Oligo (dT)18
250 µM Random Primer
dd H2O (RNase and DNase free)
1 x 20 µl
1 x 50 µl
1 x 20 µl
1 x 50 µl
1 x 20 µl
1 x 50 µl
1 x 1 ml
1 x 1 ml
–20°C (Stable for at least 12 months)
Alternatively, the solution can also be
stored at -80°C in aliquots. Avoid
repeated freezing/ thawing.
–20°C (Stable for at least 12 months)
Alternatively, the solution can also be
stored at -80°C in aliquots. Avoid
repeated freezing/ thawing.
–20°C (Stable for at least 12 months)
Alternatively, the solution can also be
stored at -80°C in aliquots. Avoid
repeated freezing/ thawing.
–20°C (Stable for at least 12 months)
Alternatively, the solution can also be
stored at -80°C in aliquots. Avoid
repeated freezing/ thawing.
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