Technical Sheet

mRNA Differential Display Kits
q 50-1000-00 Anchor-dTVN4ä
q 50-1100-00 Anchor-dTVN12ä
q 50-1200-00 Co-Anchor-GC50ä

Shipped at ambient temperature. Store at -20oC
For research use only.
 Not for use in diagnostic procedures; for clinical purposes only.

Background

Conventional Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) is a powerful tool in the amplification of rare and abundant cDNA sequences transcribed from total mRNA pool. This has been effectively utilized for mutation identification by amplifying only the target transcript of choice followed by specific sequencing. In addition this technique has been very valuable in quantitative PCR for estimating the amount of expression of a particular gene.

mRNA differential display is a form of RT-PCR in which anchored oligo dT primer is used (1). This modified oligo dT primer is designed in such a way that the last two 3’ end bases are restricted to specific combinations. The oligo dT modified primer is thus ‘anchored’ to prime only a specific subset of mRNA species which has its 3’ end before the poly A tail the cognate bases. For example, the anchored oligo dT primer; dTTTTTTTTTTTTGA will only hybridize to mRNA sequences containing TC at the 3’ end before the poly A tail. This is shown in the figure below.

mRNA
5’……TCAAAAAAAAAAAAAAAAAAAAAAAAA-3’
3’-AGTTTTTTTTTTTT-5’
(anchored oligo dT primer)

The modified anchored oligo dT primer shown above will preferentially prime the synthesis of mRNA species ending in TC at the 3’ end before the poly A tail. This technique can be used with a combination of modified primers to enable the amplification of all the transcripts from a particular developmental stage in 4 pools using Anchor-dTVN4ä (Catalog No. 50-1000-00) or in 12 pools using Anchor-dTVN12ä (Catalog No. 50-1100-00) in conjunction with Co-Anchor-GC50ä (Catalog No. 50-1200-00) to prime the arbitrary synthesis of mRNA species from the other end for PCR. Gene Link’s product Co-Anchor-GC50ä (Catalog No. 50-1200-00) is specially designed for the amplification of cDNA. It is a arbitrary sequence of a decamer with GC content of 50%. The result from PCR using the above product should yield a complex ladder pattern when electrophoretically separated on a polyacrylamide gel. Genes of interest can be comparatively identified from the gel pattern from different developmental stage or cell types and subsequently cloned by PCR (2).

References
 1. Liang,P., Averboukh, L. & Pardee, A.B. (1993) Nucleic Acids Res., 21:3269-3275.
2. Aiello, L.P., Robinson, G.S., Lin, Y-W, Nishio, Y. & King, G.L. (1994) PNAS 91:6231-6235.

Material Supplied

q 50-1000-00 Anchor-dTVN4 ä Kit $100.00

1 A260 Unit each of the following components; V=G+A+C

q50-1000-01 5’-TTT TTT TTT TTT VG-3’
q50-1000-02 5’-TTT TTT TTT TTT VT-3’
q50-1000-03 5’-TTT TTT TTT TTT VA-3’
q50-1000-04 5’-TTT TTT TTT TTT VC-3’

q 50-1100-00 Anchor-dTVN12ä Kit $250.00

1 A260 Unit each of the following components:

 q 50-1100-01 5’-TTT TTT TTT TTT GG-3’
q 50-1100-02 5’-TTT TTT TTT TTT GT-3’
q 50-1100-03 5’-TTT TTT TTT TTT GA-3’
q 50-1100-04 5’-TTT TTT TTT TTT GC-3’
q 50-1100-05 5’-TTT TTT TTT TTT AG-3’
q 50-1100-06 5’-TTT TTT TTT TTT AT-3’
q 50-1100-07 5’-TTT TTT TTT TTT AA-3’
q 50-1100-08 5’-TTT TTT TTT TTT AC-3’
q 50-1100-09 5’-TTT TTT TTT TTT CG-3’
q 50-1100-10 5’-TTT TTT TTT TTT CT-3’
q 50-1100-11 5’-TTT TTT TTT TTT CA-3’
q 50-1100-12 5’-TTT TTT TTT TTT CC-3’

q 50-1200-00 Co-Anchor-GC50ä Kit $150.00

1 A260 Unit each of the following components:

q 50-1200-01 5’-AGA AGC GAT G-3’
q 50-1200-02 5’-AGG ATA CGT G-3’
q 50-1200-03 5’-AGG TTC TAG C-3’
q 50-1200-03 5’-CAA TGC GTC T-3’
q 50-1200-04 5’-CGG ATA ACT G-3’
q 50-1200-05 5’-CCA TTT ACG C-3’
q 50-1200-06 5’-GGA AGA CAA C-3’
q 50-1200-07 5’-GTG CAA TGA G-3’
q 50-1200-08 5’-GGA AGA CAA C-3’
q 50-1200-09 5’-TCC CTT TAG C-3’
q 50-1200-10 5’-TCC GAC GTA T-3’
 
**The polymerase chain reaction (PCR) process is covered by patents owned by Hoffmann-La Roche.
A license to perform is automatically granted by the use of authorized reagents.
 All Gene Link products are for research use only. Prices subject to change without notice.

Related Products Ordering Information

Genemerä , RT-PCRmerä and LINKMERä

Genemerä , RT-PCRmerä and LINKMERä are primer pairs for specific amplification of gene, cDNA and chromosomal sequences. OligomerProbeä is available for use as hybridization probes for the above RT-PCRmerä with either 5’OH for 32P labeling or with 3’ biotin for non-radioactive detection, see OligomerProbeä section. Genemerä , RT-PCRmerä and LINKMERä primer pairs are supplied separately as lyophilized powder in aliquots of 10nmoles. The 10nmoles of primer when dissolved in 500m l sterile water or TE will give a solution of 20m Molar i.e. 20pmoles/m l. The quantity supplied is sufficient for at least 400 regular 25m l PCR reaction.

 

RT-PCRmerä *

Product
Size
Catalog No.
Price

b -actin, human
10nmoles
40-1001-10
$100.00
b -actin, rat
10nmoles
40-1002-10
$100.00
b -actin, mouse
10nmoles
40-1003-10
$100.00

b 2-microglobulin
10nmoles
40-1004-10
$100.00

G3PDH (Glyceraldehyde 3-phosphate dehydrogenase)
10nmoles
40-1005-10
$100.00

Please inquire about other RT-PCRmerä not listed here

* OligomerProbeä is available for use as hybridization probes for the above RT-PCRmerä with either 5’OH for 32P labelling or with 3’ biotin for non-radioactive detection. See OligomerProbeä section below.

 

OligomerProbeä

OligomerProbeä are specific oligonucletide probes for hybridization to its cognate species. These are specially suited for use in conjunction with RT-PCRmersä , as the complementary sequence lies in the amplified sequence. The OligomerProbeä can also be used for all northern blots. OligomerProbeä are available for use as hybridization probes with either 5’OH for 32P labeling or with 3’ biotin for non-radioactive detection. The OligomerProbeä is supplied as a lyophilized powder in aliquots of 2nmoles. The 2nmoles of primer when dissolved in 100m l sterile water or TE will give a solution of 20m Molar i.e. 20pmoles/m l.

 

OligomerProbeä

Product
Size
Catalog No.
Price

b -actin, human
2nmoles
40-1101-02
$55.00

b -actin, rat
2nmoles
40-1102-02
55.00

b -actin, mouse

2nmoles
40-1103-02
$55.00

b 2-microglobulin
2nmoles
40-1104-02
$55.00

G3PDH (Glyceraldehyde 3-phosphate dehydrogenase)
2nmoles
40-1105-02
$55.00

Please inquire about other OligomerProbeä not listed here

 

 

GENEMERä

Product
Size
Catalog No.
Price

Sickle Cell SC2/SC5 primer pair
10nmoles
40-2001-10
$100.00

RhD (Rh D gene exon 10 specific)
10nmoles
40-2002-10
$100.00

Rh EeCc (Rh Ee and Cc exon 7 specific)
10nmoles
40-2003-10
$100.00

Fragile X (spanning triple repeat region)

10nmoles
40-2004-10
$100.00

Gaucher 1226G mutation specific
10nmoles
40-2005-10
$100.00

Gaucher 1448C mutation specific
10nmoles
40-2006-10
$100.00

Gaucher 84GG mutation specific
10nmoles
40-2007-10
$100.00

Gaucher IVS2 mutation specific
10nmoles
40-2008-10
$100.00

Cystic Fibrosis D F508

10nmoles
40-2009-10
$100.00

Cystic Fibrosis G542X
10nmoles
40-2010-10
$100.00

Cystic Fibrosis W1282X
10nmoles
40-2011-10
$100.00

Cystic Fibrosis G551D/R553X
10nmoles
40-2012-10
$100.00

Cystic Fibrosis N1303K
10nmoles
40-2013-10
$100.00

Cystic FibrosisCT3849
10nmoles
40-2014-10
$100.00

Please inquire about other GENEMERä not listed here

 

LINKMERä

LINKMERä are primer pairs for specific amplification of chromosomal sequences; specially suited for linkage analysis and chromosomal mapping. LINKMERä are supplied as lyophilized powder in aliquots of 10nmoles. The 10nmoles of primer when dissolved in 500m l sterile water or TE will give a solution of 20m Molar i.e. 20pmoles/m l. The quantity supplied is sufficient for at least 400 regular 25m l PCR reaction.

 

Product
Size
Catalog No.
Price

Please inquire, Linkmersä

10nmoles
  
$75.00

 

**The polymerase chain reaction (PCR) process is covered by patents owned by Hoffmann-La Roche. A license to perform is automatically granted by the use of authorized reagents.



Prices subject to change without notice
All Gene Link products are for research use only.

Please direct all pricing inquiries to our Orders department.
For any other problems, please contact our WebMaster.