FastLane Cell cDNA Kit

For high-speed preparation of cDNA without RNA purification for use in real-time RT-PCR
  • From cells to cDNA in only 4 steps and under 45 minutes
  • Sensitive detection of even low-abundance transcripts
  • Easy parallel processing of several samples
  • Integrated gDNA removal allows detection of RNA only
  • No need to design RNA-specific primers or probes

The FastLane Cell cDNA Kit provides a fast and simple procedure for preparing first-strand cDNA directly from cultured cells without RNA purification. The kit is supplied with wash and lysis buffers for preparing lysates and stabilizing RNA, gDNA Wipeout Buffer for eliminating genomic DNA contamination, and all reaction components for fast and efficient cDNA synthesis. The FastLane Cell cDNA Kit is ideal for experiments requiring rapid, high-throughput gene expression analysis.

产品 Product no. 货号 目录价:
 
 
Show details
    varies
Can't order online?
To place an order via phone, email or for requesting a quote, please provide the product’s name, number and catalog number.
产品 货号 目录价:
FastLane Cell cDNA Kit (50)
Buffer FCW, Buffer FCP, and components for 50 x 20 µl reverse-transcription reactions (gDNA Wipeout Buffer, Quantiscript Reverse Transcriptase, Quantiscript RT Buffer, RT Primer Mix, and RNase-Free Water)
215011
询价

FastLane Cell cDNA Kit适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。


  • Main Image Navi
出色的灵敏度适用于癌症研究。|成功分析CDC2敲除。|高效去除基因组DNA。|高度可重复性的制备cDNA。|
在HeLa细胞中对重要的癌症研究基因进行两步法real-time RT-PCR分析时,使用FastLane Kit制备cDNA获得的CT值低于使用Supplier AIV的同类试剂盒制备cDNA获得的CT值。|使用不同的CDC2 siRNA(、S2S3)或非沉默siRNA(NS)转染MCF-7细胞。48小时后,使用FastLane Kit制备cDNA。利用两步法real-time RT-PCR分析CDC2(胞周期蛋白)相对于GAPDH(内源性对照)的表达。将未处理细胞(UT)中CDC2的相对表达水平设置为100%。|使用FastLane Cell cDNA Kit,加入(+RT)或不加入(–RT)逆转录酶,制备cDNA。使用针对β-actin的基因表达分析定量cDNA。-RT曲线显示未检测到基因组DNA。|使用FastLane Kit,在包含HeLa细胞的48孔板中制备cDNA。利用两步法real-time RT-PCR分析微管蛋白B(结构蛋白)的表达。|
性能
cDNA synthesized by the FastLane Cell cDNA Kit enables highly sensitive and reproducible results in real-time two-step RT-PCR (see figures "Superior sensitivity in cancer research" and "Highly reproducible cDNA preparation"). The high quality of the cDNA also allows detection over a wide linear range from >105 cells down to one cell.
原理

The FastLane Cell cDNA Kit uses FastLane technology to prepare cDNA directly from cultured cells in just 45 minutes. To ensure only RNA is detected in real-time RT-PCR, novel gDNA Wipeout Buffer is used to eliminate genomic DNA (see figure "Effective elimination of genomic DNA contamination"). Time and effort are saved as there is no need to design RNA-specific primers or probes.

When the FastLane Cell cDNA Kit is used together with optimized QuantiTect, QuantiFast, or Rotor-Gene Kits for real-time RT-PCR, several cell samples can be easily processed and analyzed within a few hours. This allows a snapshot of several transcripts in experiments such as validation of siRNA-mediated gene knockdown (see figure "Successful analysis of CDC2 knockdown").

The FastLane Cell cDNA Kit is part of the comprehensive range of FastLane Kits, which speed up and streamline the gene expression analysis workflow.

操作流程

With the FastLane Cell cDNA Kit, cDNA is obtained from cultured cells in just 45 minutes in a 4-step procedure: cell wash; cell lysis with integrated RNA stabilization; genomic DNA elimination; and cDNA synthesis. The synthesized cDNA is ready to use in real-time two-step RT-PCR.

The kit contains sufficient reagents for preparing cDNA from two 24-well cell-culture plates or one 48-well cell-culture plate. If working with a 96-well cell-culture plate, an additional QuantiTect Reverse Transcription Kit (50) needs to be purchased.

cDNA prepared with the FastLane Cell cDNA Kit can be analyzed by real-time two-step RT-PCR using one of the following QuantiTect Kits:

  • SYBR® Green detection: QuantiTect SYBR® Green PCR Kit
  • Probe detection: QuantiTect Probe PCR Kit
  • Multiplex probe detection: QuantiTect Multiplex PCR Kits

Alternatively, fast real-time two-step RT-PCR can be carried out when the FastLane Cell cDNA Kit is used in combination with a QuantiFast Kit:

  • SYBR® Green detection: QuantiFast SYBR® Green PCR Kit
  • Probe detection: QuantiFast Probe PCR Kits
  • Multiplex probe detection: QuantiFast Multiplex PCR Kits

Ultrafast real-time two-step RT-PCR can be achieved when cDNA prepared with the FastLane Cell cDNA Kit is analyzed on the Rotor-Gene Q cycler using one of the following dedicated Rotor-Gene Kits:

  • SYBR® Green detection: Rotor-Gene SYBR® Green PCR Kit
  • Probe detection: Rotor-Gene Probe PCR Kit
  • Multiplex probe detection: Rotor-Gene Multiplex PCR Kit 

QuantiTect and QuantiFast Kits are compatible with various real-time cyclers, including instruments from Applied Biosystems.

应用

The FastLane Cell cDNA Kit is suitable for experiments which require a snapshot of individual transcript levels, such as:

  • Validation of siRNA-mediated gene knockdown
  • Evaluation of drug effects
  • Detection of gene regulation
特点
参数
应用 Real-time, two-step RT-PCR, gene expression analysis directly from cells
酶活 Reverse transcription
预混液 No
反应类型 cDNA production, DNA digestion
real-time或终点法PCR Real-time, two-step RT-PCR
样本/目标类型 Cultured cells
单一或多重 Single
有/无热启动酶 Without hotstart

您无权限查看此资源

Kit Handbooks
1
For high-speed preparation of first-strand cDNA directly from cultured cells without RNA purification. For use in real-time, two-step RT-PCR
显示更多
图片
Superior sensitivity in cancer research
出色的灵敏度适用于癌症研究。
在HeLa细胞中对重要的癌症研究基因进行两步法real-time RT-PCR分析时,使用FastLane Kit制备cDNA获得的CT值低于使用Supplier AIV的同类试剂盒制备cDNA获得的CT值。
Successful analysis of CDC2 knockdown
成功分析CDC2敲除。
使用不同的CDC2 siRNA(、S2S3)或非沉默siRNA(NS)转染MCF-7细胞。48小时后,使用FastLane Kit制备cDNA。利用两步法real-time RT-PCR分析CDC2(胞周期蛋白)相对于GAPDH(内源性对照)的表达。将未处理细胞(UT)中CDC2的相对表达水平设置为100%。
Effective elimination of genomic DNA contamination
高效去除基因组DNA。
使用FastLane Cell cDNA Kit,加入(+RT)或不加入(–RT)逆转录酶,制备cDNA。使用针对β-actin的基因表达分析定量cDNA。-RT曲线显示未检测到基因组DNA。
Highly reproducible cDNA preparation
高度可重复性的制备cDNA。
使用FastLane Kit,在包含HeLa细胞的48孔板中制备cDNA。利用两步法real-time RT-PCR分析微管蛋白B(结构蛋白)的表达。