CALL (518)-605-8808 or
FAX: (206)-337-1074 FOR ORDERING
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
CALL (518)-605-8808 or
FAX: (206)-337-1074 FOR ORDERING
|
