SuperPrep® II cell Lysis & RT Kit for qPCR, Trial (Only 1 Per Customer)
SuperPrep® II cell Lysis & RT Kit for qPCR (*Only 1 Per Customer*)
Cat. No. SCQ-401
Store -20ºC
Description
"Lysis Reagents" prepares cell lysates containing RNAs that can be used as templates for reverse transcription. "RT Reagents" contains reagents for reverse transcription, optimized for efficient cDNA synthesis from crude lysates. The synthesized cDNA can be applied to real-time PCR directly. This assay system is suitable for high-throughput assays. In the previous version, it was necessary to add Stop Solution after adding Lysis Solution. In SuperPrep® II, cell lysates with lysis solution can be used as templates for cDNA synthesis directly. Moreover, high sensitivity detection is possible from a wide variety of mammalian cells than in the previous version. With this assay system, it is possible to synthesize template cDNA for real-time PCR from cultured cells conveniently and quickly.
SuperPrep® II Cell Lysis Kit for qPCR (Code No. SCQ-501) is an option of "Lysis Reagents". The cell lysate prepared by "Lysis Reagents" can be applied to one-step real-time PCR.
- RNA purification is not necessary.
- High-quality cDNA can be obtained from cell lysates.
- Reduction of dispersion on high-throughput assay.
- Various real-time PCR regents can be applied.
Total RNA preparation for one-step real-time PCR from mammalian cultured cells (Code No. SCQ-501)
Components
SCQ-401S and SCQ-401 contain two separate packages named "Lysis Reagents" and "RT Reagents", respectively. All reagents should be stored at -20°C
Table 1.SuperPrep® II Cell Lysis & RT Kit for qPCR (Code No.SCQ-401, SCQ-401S)
|
<Lysis Reagents> |
TYB-SCQ-401 |
TYB-SCQ-401S |
|
Lysis Solution |
6.5 ml |
1.3 ml |
|
gDNA Remover |
33 µl |
6.6 µl |
|
RNase Inhibitor |
110 µl |
22 µl |
| Link |  |
|
|
<RT Reagents> |
||
|
5 × RT Master Mix |
860 µl |
172 µl |
|
5 × RT Master Mix no-RT Control |
86 µl |
17 µl |
|
Nuclease-free Water |
1.7 ml × 2 |
680 µl |
Table 2.SuperPrep® II Cell Lysis Kit for qPCR (Code No.SCQ-501)
|
Lysis Solution |
6.5 ml |
|
gDNA Remover |
33 µl |
|
RNase Inhibitor |
110 µl |
Applications
The optimized lysis solution efficiently inhibits RNA degradation during treatment. RNA in the lysate is stable on ice for at least 6 h. High-quality cDNA can be synthesized using highly efficient reverse transcriptase "ReverTra Ace®" with low contamination of genomic DNA because of preceding DNase I treatment. The reverse transcriptase is supplied as a master mix reagent containing optimally mixed primers (random and oligo dT) to achieve effective cDNA synthesis.
Table 1 Cells tested by this system
|
Call Name |
Adherent |
Species |
Remarks |
|
|
1 |
HPA |
Adherent |
H.sapiens |
preadipocytes (primary cell) |
|
2 |
HEK |
Adherent |
H.sapiens |
epidermal keratinocytes (primary cell) |
|
3 |
HA |
Adherent |
H.sapiens |
astrocytes (primary cell) |
|
4 |
HDF |
Adherent |
C. griseus |
dermal fibroblasts (primary cell) |
|
5 |
HBEpC |
Adherent |
H.sapiens |
bronchial epithelial cells (primary cell) |
|
6 |
HUVEC |
Adherent |
H.sapiens |
umbilical vein endothelial cells (primary cell) |
|
7 |
HPAEC |
Adherent |
H.sapiens |
pulmonary artery endothelial cells (primary cell) |
|
8 |
HC |
Adherent |
H.sapiens |
chondrocytes (primary cell) |
|
9 |
HOb |
Adherent |
H.sapiens |
osteoblasts (primary cell) |
|
10 |
HSkMC |
Adherent |
H.sapiens |
skeletal muscle cells (primary cell) |
|
11 |
HAOSMC |
Adherent |
H.sapiens |
Aortic smooth muscle cells (primary cell) |
|
12 |
HFDPC |
Adherent |
H.sapiens |
hair follicle dermal papilla Cells (primary cell) |
|
13 |
HeLa S3 |
Adherent |
H.sapiens |
cervix carcinoma cell line |
|
14 |
HepG2 |
Adherent |
H.sapiens |
hepatocellular carcinoma cell line |
|
15 |
Jurkat |
Non-adherent |
H.sapiens |
T lymphocyte cell line |
|
16 |
K562 |
Non-adherent |
H.sapiens |
myelogenous leukemia cell line |
|
17 |
THP-1 |
Non-adherent |
H.sapiens |
acute monocytic leukemia cell line |
|
18 |
U937 |
Non-adherent |
H.sapiens |
leukemic monocyte lymphoma cell line |
|
19 |
HMNC |
Non-adherent |
H.sapiens |
Mononuclar cells (primary cell) |
RNase activity depends on the type and number of cells. The cell lysates should be placed on ice after preparation and cDNA should be synthesized immediately after preparing the lysates to minimize RNA degradation.
Example 3.Various real-time PCR reagents can be applied.
Z’ factors from SuperPrep® were superior to that from the other system (Company A).
*The Z’ factor is a simple statistical parameter that is used to assess the quality of high-throughput screening (HTS) assays. A Z’ score of ≥0.5 is generally considered to indicate good quality Z’ can be calculated by the following formula.
Z’= 1-3 x [Δ Ct(+) standard deviation + Δ Ct(-) standard deviation]/| Δ Δ Ct|
