Invitrogen™ GeneRacer™ Kit with SuperScript™ III RT and TOPO TA Cloning™ Kit for Sequencing

Ensures amplification of only full-length transcripts via elimination of truncated messages from amplification process

Supplier:  Invitrogen™ L150201

Catalog No. L150201

Description

The GeneRacer Kit provides a method to obtain full-length 5' and 3' ends of cDNA using known cDNA sequence from expressed sequence tags (ESTs), subtracted cDNA, differential display, or library screening. RACE PCR products can be quickly and easily cloned using either the Zero Blunt TOPO PCR Cloning Kit for Sequencing (blunt-end PCR products) or the TOPO TA Cloning for Sequencing Kit (PCR products with 3' A-overhangs). Using the protocols provided, the cDNA ends of rare (30 copies/cell) and long (9 kb) transcripts can be amplified and sequenced starting from 1μg of total RNA.

• Generate cDNA from transcripts at least 10kb in length
• Obtain full-length 5' end of rare transcripts at fewer than 30 copies per cell
• Clone full-length 5' and 3' ends to construct complete cDNA sequence

Cloning, PCR and Real-Time PCR, Reverse Transcription, cDNA Libraries and Library Construction

Order Info

Shipping Condition: Dry ice

Specifications

• Bacterial or Yeast Strain: TOP10
• Content And Storage: Store each module as indicated:
  
  GeneRacer™ Module (-20°C):
  
  • 2 × 1.5 ml Sterile Water
  
  • 24 μl RNaseOut™
  
  • 6 μl each Calf Intestinal Phosphatase (CIP), CIP Buffer, Tobacco Acid Pyrophosphatase (TAP), 10X TAP Buffer, T4 RNA Ligase, 10X T4 RNA Ligase Buffer, and 10 mM ATP
  
  • 6 × 250 ng GeneRacer™ RNA Oligo
  
  • 2 × 1 ml Phenol/Chloroform
  
  • 36 μl Mussel Glycogen
  
  • 200 μl Sodium Acetate (3 M)
  
  • 225 μl each GeneRacer™ 5′ Primer, 5′ Nested Primer, 3′ Primer, and 3′ Nested Primer
  
  • 20 μl Control HeLa Total RNA (500 ng/μl)
  
  • 15 μl each Control Primer A and Control Primer B.1
  
  SuperScript™ III RT Module (-20°C)
  
  • 6 μl SuperScript™ III Reverse Transcriptase (200 U/μl)
  
  • 24 μl 5X First Strand Buffer
  
  • 15 μl DTT (0.1 M)
  
  • 6 μl RNaseH (2 U/μl)
  
  • 6 μl Random Primers (100 ng/μl)
  
  • 6 μl GeneRacer™ Oligo dT Primer (900 ng/μl)
  
  • 6 μl dNTP Mix (10 mM each)
  
  10 S.N.A.P.™ Columns (room temperature)
  
  TOPO TA Cloning™ Kit for Sequencing (-20°C)
  
  • Sufficient reagents and One Shot™ TOP10 Chemically Competent E. coli (store at -80°C) to clone 10 GeneRacer™ PCR products; GeneRacer Module (-20°C), SuperScript III RT Module (-20°C), Competent E. coli (store at -80°C), S.N.A.P. Columns (room temperature), TOPO TA Cloning Kit for Sequencing (-20°C)
• For Use With (Application): Reverse Transcription
• Product Line: GeneRacer™, SuperScript™, TA Cloning™, TOPO™
• Quantity: 1 Kit

• Cloning Method: TOPO TA
• Format: Kit
• Includes: GeneRacer Module: 2 x 1.5mL Sterile Water, 24μL RNaseOut, 6μL each Calf Intestinal Phosphatase (CIP), CIP Buffer, Tobacco Acid Pyrophosphatase (TAP), 10X TAP Buffer, T4 RNA Ligase, 10X T4 RNA Ligase Buffer, and 10mM ATP, 6 x 250ng GeneRacer RNA Oligo, 2 x 1mL Phenol/Chloroform, 36μL Mussel Glycogen, 200μL Sodium Acetate (3M), 225μL each GeneRacer 5' Primer, 5' Nested Primer, 3' Primer, and 3' Nested Primer, 20μL Control HeLa Total RNA (500ng/μL), 15μL each Control Primer A and Control Primer B.1; SuperScript III RT Module: 6μL SuperScript III Reverse Transcriptase (200U/μL), 24μL 5X First Strand Buffer, 15μL DTT (0.1M), 6μL RNaseH (2U/μL), 6μL Random Primers (100ng/μL), 6μL GeneRacer Oligo dT Primer (900ng/μL), 6μL dNTP Mix (10mM each), 10 S.N.A.P. Columns, TOPO TA Cloning Kit for Sequencing (-20°C), Sufficient reagents and One Shot TOP10 Chemically Competent E. coli
• Product Type: Cloning Kit
• Vector: pCR4-TOPO TA

For Research Use Only. Not for use in diagnostic procedures.