Product FAQ

Can the compatibility reagent in the BCA-RACE kit be stored after reconstitution and reused?

Answer

The reconstituted compatibility reagent is stable for 8 hours when stored at 4 degrees C protected from light.

Answer Id: E8306

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Product FAQ

I’m getting ambiguous results with my FirstChoice RLM-RACE Kit. Do you have any suggestions?

Answer

Without optimization, nested PCR may produce no band, a single band, several bands, or a complicated pattern of bands (a smear). Smearing or failure to amplify could alternatively be caused by poor-quality RNA, or absence of the target in the RNA used for RLM-RACE. Ensure that pure, high-quality RNA is being used as your starting material.

-Multiple bands can be due to multiple initiation sites for transcription of the target gene or primer homology; try a new set of primers. If you are using inner and outer gene specific 5’ RLM RACE primers (for nested PCR), try using each one of them in a PCR with a gene specific 5’ primer and an aliquot of the RLM-RACE RT reaction as template.
-Perform the minus-TAP control; the minus-TAP control RNA should not yield the same PCR products as the experimental RNA you are using. No bands should be produced since RNA has either been dephosphorylated with CIP or it has an intact cap structure that can’t undergo ligation to the 5’ RACE Adapter.
-Optimize the PCR annealing temperature; the annealing temperature of the outer PCR is less critical and should be 55-65 degrees C while the inner, nested PCR may need to be higher to achieve the required selectivity in the amplification. If the PCR fails to give the expected results, repeat the experiment using a higher (try 2 degrees C higher) annealing temperature.
-If you have a longer target, the amplification cycle can be extended 1 min for each kilobase of target over 1kb.
-If you have a high GC-rich region and/or other regions of stable secondary structure in the RNA transcripts, you can try increasing the temperature of the RT reaction so that secondary structure effects can be minimized. The M-MLV RT included can be used at up to 50 degrees C. Increasing the temperature of the synthesis reaction may facilitate read-through by the RT enzyme.

Answer Id: E7336

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Manual / Product Insert

Protocol: FirstChoice® RLM-RACE Kit (English )

Version: 09 Sep 2010
Catalog #
  • AM1700
  • AM1700M
  • AM2050(Discontinued)
  • AM2052(Discontinued)
  • AM2054(Discontinued)
  • AM2056(Discontinued)

Manual / Product Insert

FirstChoice® RLM-RACE Kit Instruction Manual

Version: 0803C
Catalog #

Citations & References

Molecular cloning of a novel chaperone-like protein induced by rhabdovirus infection with sequence similarity to the bacterial extracellular solute-binding protein family 5.

  • Authors: Cho Wha Ja; Yoon Won Joon; Moon Chang Hoon; Cha Seung Ju; Song Hebok; Cho Hong Rae; Jang Soo Jin; Chung Dae Kyun; Jeong Choon Soo; Park Jeong Woo;
  • Journal: J Biol Chem (2002) 277:41489-41585
  • PubMed ID: 12183466

Citations & References

Mediation of the DCC apoptotic signal by DIP13 alpha.

  • Authors: Liu Jiayou; Yao Fayi; Wu Ruping; Morgan Michael; Thorburn Andrew; Finley Russell L Jr; Chen Yong Q;
  • Journal: J Biol Chem (2002) 277:26281-26286
  • PubMed ID: 12011067
Catalog #

Citations & References

Characterization of the 5' regulatory region of the human sodium-dependent multivitamin transporter, hSMVT.

  • Authors: Dey S, Subramanian VS, Chatterjee NS, Rubin SA, Said HM,
  • Journal: Biochim Biophys Acta (2002) 1574:187-192
  • PubMed ID: 11955628
Catalog #

Product Literature

Bangalore Warehouse stock

Citations & References

Characterization of Drosophila hemoglobin. Evidence for hemoglobin-mediated respiration in insects.

  • Authors: Hankeln Thomas; Jaenicke Viviane; Kiger Laurent; Dewilde Sylvia; Ungerechts Guy; Schmidt Marc; Urban Joachim; Marden Michael C; Moens Luc; Burmester Thorsten;
  • Journal: J Biol Chem (2002) 277:29012-29019
  • PubMed ID: 12048208
Catalog #

Product Literature

GeneRacer RLM-Racer Kit

Citations & References

Myeloid expression of cytochrome P450 4F3 is determined by a lineage-specific alternative promoter.

  • Authors: Christmas P, Carlesso N, Shang H, Cheng SM, Weber BM, Preffer FI, Scadden DT, Soberman RJ,
  • Journal: J Biol Chem (2003) 278:25133-25142
  • PubMed ID: 12709424
Catalog #
  • K131002(Discontinued)
  • D18374025

Manual / Product Insert

FirstChoice® RACE-Ready cDNA Instruction Manual

Version: 0803C

Citations & References

Cloning of a novel retinoid-inducible serine carboxypeptidase from vascular smooth muscle cells.

  • Authors: Chen J; Streb J W; Maltby K M; Kitchen C M; Miano J M;
  • Journal: J Biol Chem (2001) 276:34175-34256
  • PubMed ID: 11447226

Citations & References

A biochemical and molecular characterization of LEP1, an extensin peroxidase from lupin.

  • Authors: Price NJ, Pinheiro C, Soares CM, Ashford DA, Ricardo CP, Jackson PA,
  • Journal: J Biol Chem (2003) 278:41389-41399
  • PubMed ID: 12882982
Catalog #

Citations & References

Primary structure and characteristics of a lectin from skin mucus of the Japanese eel Anguilla japonica.

  • Authors: Tasumi Satoshi; Ohira Tsuyoshi; Kawazoe Ichiro; Suetake Hiroaki; Suzuki Yuzuru; Aida Katsumi;
  • Journal: J Biol Chem (2002) 277:27305-27316
  • PubMed ID: 11959866
Catalog #
  • 15620016(Discontinued)