タイトルまたは説明
Product name:
First-strand cDNA Synthesis Mix
(third generation enzyme, independent primers)
Cat. No.: F***2O
Price: $**0 for *0 ul***0T(negotiable)
Storage conditions: **0°C
Product components:
|
components
|
specification
|
|
RTase III Primer
Flexible All-in-One Mix
|
**0μl
|
|
Oligo(dT)*0VN
(*0μM)
|
**0μl
|
|
Random hexamers
(*0μM)
|
**0μl
|
|
Nuclease-Free
Water
|
2×1
ml
|
Product Introduction
RTase III Primer Flexible All-in-One Mix is
​​a high-efficiency,
low-pollution, high-quality, single-strand cDNA synthesis premix.
It contains M-MLV GIII Reverse Transcriptase and its reaction
buffer, RNase inhibitors, dNTPs and other components required for
single-strand cDNA synthesis. You only need to add RNA template,
primers and water to start the reaction. It is very convenient to
operate. Different types of reverse transcription primers can be
used flexibly for different experimental designs to meet diverse
experimental needs. Oligo(dT)*0VN or Random hexamers or Gene
Specific Primers can be selected according to different
experimental scenarios. Using this reverse transcription premix, a
maximum of *2 kb cDNA can be obtained within *5 minutes.
Instructions
Prepare the following reaction system on ice:
|
reagents
|
Amount of usage
|
|
Template
RNAa
|
*0
ng~1μg
|
|
RTase III Primer
Flexible All-in-One Mix
|
4μl
|
|
Oligo(dT)*0VN
(*0μM)
|
1μl
|
|
or Random hexamers
(*0μM)
|
1μl
|
|
or Gene
Speciï¬c
Primers (*0μM)
|
0.1μl
|
|
Nuclease-Free
Water
|
To
*0μl
|
a. It is recommended to use high-quality RNA extracted by the kit
and freed of genomic DNA contamination as a template.
2. Gently pipette to mix and spin;
3. Incubate at *5℃ for *5 min;
Note: If the target RNA does not contain a Poly(A) structure, it
can be pre-incubated at *5℃ for *0 min.
4. After the reaction, incubate at *5℃ for 5 min to terminate the
reaction;
5. Place the obtained cDNA solution on ice for subsequent
experiments; or store it immediately at **0℃.
Note:
1. The subsequent experiment is a cloning experiment. If the RNA
comes from eukaryotes, only Oligo (dT) VN is needed. Adding Random
hexamers will reduce the yield of full-length cDNA; if the RNA
comes from prokaryotes, only Random hexamers or Gene
Speci
ï¬c Primers are
needed.
2. The subsequent experiment is qPCR. Please add Oligo(dT) VN and
Random hexamers at the same time to obtain cDNA with uniform
reverse transcription efficiency at different positions of
mRNA.
Notes
To prevent RNase contamination, please keep the experimental area
clean; wear clean gloves and masks during operation; the
consumables such as centrifuge tubes and gun tips used in the
experiment must be RNasefree.
