Molecular Biology Products


PCR Primer Sets

The single most important component of a successful PCR project is a set of carefully designed oligonucleotide primers. Several factors go into making successful PCR primers including; G+C content, target secondary structure, primer length, inter- and intra- primer complementarity, melting temperature and stability. We have taken all the trouble out of primer design by doing it for you! Each primer pair has been optimized for use in reverse transcriptase PCR techniques. Complete instructions for PCR conditions and 10X PCR buffer is included.

 

PCR Primer Pairs 100 pmol/m l, 50 m l – $100/each

(Enough for 500 PCR reactions. Includes instructions and 10X PCR buffer)

10X PCR Buffer, $25/1 ml

 

Housekeeping Genes

Rat, Mouse or Human b -actin

Rat GAPDH

 

Lipid Metabolism

Rat, or Mouse acyl-CoA oxidase (ACO)

Rat L-Fatty Acid Binding Protein (FABP)

Rat Acyl-CoA synthetase (ACS)

Rat acyl-CoA binding protein (ACBP)

Rat or Mouse peroxisome proliferator-activated receptor a (PPARa )

Mouse peroxisome proliferator-activated receptor b (PPARb )

Mouse peroxisome proliferator-activated receptor g (PPARg , g 1 or g 2 specific)

Mouse adipocyte protein-2 (mAP2)

 

Xenobiotic-Inducible

Human, rat, mouse, lake trout Cytochrome P450 1A1 (CYP1A1)

Rat Cytochrome P450 1A2 (CYP1A2)

Rat Cytochrome P450 IVA1 (CYP4A1)

Rat UDP-glucoronyl transferase 1 (UGT1)

Rat c-myc

Rat Proliferating cell nuclear antigen (PCNA)

 

 

PCR Internal Standards

 

The internal standard (IS) is a key reagent in the quantitation of mRNA or DNA using PCR. The requirement of an internal standard is necessitated by the fact that there is a large amount of tube-to-tube variability in amplification efficiency. For example if 10 tubes of seemingly identical reagents are PCR amplified, there could be as much as a 3-fold difference in the amount of product formed. If an IS was co-amplified with the target, the efficiency of amplification in each tube could be corrected and this 3-fold difference could easily be negated. However, not all internal standards are created equally. One should keep in mind the requirements of their internal standard, i.e. what criteria is being applied to assess whether an IS is adequate. Some of the major criteria are listed below:

An internal standard should amplify with the same efficiency as the target. This is usually accomplished by incorporating the target gene’s primer sequences in the IS and making it of approximately the same length as the target PCR product.

The product from the IS must be easy to resolve from the target. This may be accomplished by making the two products different enough in length to be resolved on a gel or by adding a restriction enzyme recognition site.

The internal standard should control for differences in reverse transcription efficiency. This is can be done by making the IS an RNA molecule with a poly(A)+ tail.

Our internal standards meet all these requirements and have been tested and optimized. Recombinant RNA molecules are synthesized as described by Vanden Heuvel et al., and shipped on dry ice, quantitated and ready for use in competitive RT-PCR.

 

rcRNA internal standards 10 ng/m l, 20 m l $250

(Shipped on dry ice. Rnase free. Included with 10X PCR buffer and primers necessary for 250 PCR reactions and complete instructions).

 

Housekeeping Genes

Rat, Mouse or Human b -actin

Rat GAPDH

 

Lipid Metabolism

Rat, or Mouse acyl-CoA oxidase (ACO)

Rat L-Fatty Acid Binding Protein (FABP)

Rat or Mouse peroxisome proliferator-activated receptor a (PPARa )

Mouse peroxisome proliferator-activated receptor b (PPARb )

Mouse peroxisome proliferator-activated receptor g (PPARg )

Mouse adipocyte protein-2 (mAP2)

 

Xenobiotic-Inducible

Human, rat, mouse, lake trout Cytochrome P450 1A1 (CYP1A1)

Rat Cytochrome P450 1A2 (CYP1A2)

Rat Cytochrome P450 IVA1 (CYP4A1)

Rat UDP-glucoronyl transferase 1 (UGT1)

Rat c-myc

Rat Proliferating cell nuclear antigen (PCNA)

 

 

Custom PCR Internal Standards

If we do not have the primers or internal standard necessary for your project, we can design and synthesize the necessary reagents required to perform quantitative PCR. You provide us the information on the gene to be studied and we will do the rest, including optimizing the primer sets. Please specify whether a recombinant RNA (rcRNA) or DNA internal standard is required.

 

Custom Internal Standards

DNA 10 ng/m l, 20 m l $250

(Shipped on wet ice. Included with 10X PCR buffer and primers necessary for 250 PCR reactions and complete instructions).

rcRNA 10 ng/m l, 20 m l $500

(Shipped on dry ice. Rnase free. Included with 10X PCR buffer and primers necessary for 250 PCR reactions and complete instructions).

 

Quantitative PCR Services

From training to RNA or DNA extraction, primer design and specific gene quantitation, we can meet any or all of your quantitative PCR needs. Use our experience and get the data that your project requires – without the headaches.

 

Quantitative PCR Services Please Inquire

Training

Primer design and optimization

DNA or RNA extraction

Internal standard design and synthesis

Quantitation of specific gene

Quantitation of housekeeping gene

Data analysis