DDRT-PCR Kit
A Differential Display Kit from Display Systems Biotech

One of the most convenient methods of determining the effect of a given chemical or environmental influence on the expression of genes is "differential display". The Differential Display (DDRT-PCR) Kit from Display Systems Biotech is a comprehensive and widely used system for identifying and isolating genes that are up or down regulated. The DDRT-PCR system allows the researcher to generate a "snapshot" of the "before" and "after" expression profile of a cell. The procedure provides for direct cloning of any differences observed and is thus a rapid and powerful means of characterizing modulation in gene expression

The basic protocol for the DDRT-PCR Kit is as follows:

1. Total RNA is isolated from control and test cells and reverse transcribed using a set of localized primers. Each primer pro- duces a cDNA population that represents an overlapping sub- set of the total expression profile of the cell.
2. Each subset cDNA population is PCR-amplified using the original primer and a number of specific proprietary primers in the prescence of 35S or 33P dATP.
3. Amplified products from control and test cells are separated by gel electrophoresis and band intensity differences are used to identify expression-modulated mRNA species. Observed differences are confirmed by SSCP and RPA analysis.
4. Differentially displayed bands are excised from the gel, reamplified , cloned (optional), and identified by sequence analysis.

DDRT-PCR Scheme

DDRT-PCR Kit from Display Systems
Cat. No.	Description	Size/Vol.
DSB500-100	Differential Display Kit	4 x 96 rxns*
Components
DSB500-101:124	Upstr. primer 1-24	500 痞/2 然
DSB500-125:133	Downstr. primer 1-9	500 痞/25 然
DSB500-134	dNTP, 0.5 ml x 2	500 然
DSB500-135	Magnesium Chloride	2 ml/25 mM
DSB500-136	T7 Sequencing Primer	100 痞/2 然
DSB500-137	Ext. primer T7-T11-A	100 痞/2 然
DSB500-138	Ext. primer T7-T11-C	100 痞/2 然
DSB500-139	Ext. primer T7-T11-G	100 痞/2 然
DSB551-100	displayTAQ ML	100 痞/5u/痞
DSB500-141	displayTAQ 10X Buf.	2 ml
DSB500-142	RT, MMuLV, 20 u/痞	12.5 痞
DSB500-143	RT, 5X First-strand Buf.	2 ml
DSB500-146	cycleSEQ, man., enz	20 templates
DSB500-147	cycleSEQ buffer
DSB500-148	cycleMix A
DSB500-149	cycleMix C
DSB500-150	cycleMix G
DSB500-151	cycleMix T
DSB500-152	cycleSEQ control DNA
*Enough displayTAQ ML for 4 x 96 rxns; enough RT, MMuLV for 2 x 96 rxns.
Shipped on dry ice; store frozen at -20°C.
Also Available:
DSB500-240	displayTAQ, 1000 u	200 痞, 5 u/痞

displayENZYME Kit
DSB520-100	displayENZYME Kit
DSB551-200	displayTAQ ML, 1000 u	200 痞, 5 u/痞
DSB500-241	displayTAQ, 10X Buf.	4 ml
DSB500-242	RT MMuLV, 500 u	25 痞,20 u/痞
DSB500-243	RT 5X 1st-strand Buffer	1 ml
Shipped on dry ice; store frozen at -20°C.