Introducing our latest panel: Mouse Thymus!

(See a complete list of available Rapid-Screen™ tissue-specific libraries

and catalog numbers at the bottom of this page)

Advantages RAPID Only 3 sets of PCR reactions are needed to idenify desired cDNA clones. Proceed quickly from the beginning of your PCR screen to cell culture transfection with your positive clones. POWERFUL cDNAs as large as 10 kb. 500,000 clones in each library. EASY No colony screening by DNA hybridization needed. No large scale plating of bacteria and no pulling of filters needed. Both degenerate and specific primers can be used to screen these panels. Libraries are ligated into a mammalian expression vector with the CMV promoter, so your clones are ready for expression in tissue culture cells. Overview The Rapid-Screen cDNA Library Panels are an innovative cloning system for the easy and rapid identification of cDNA clones. Each cDNA library panel is arrayed so that you will be able to identify your positive clone of intererst with as few as 3 sets of PCR reactions. These libraries are made in the mammalian expression vector pCMV6-XL4. This vector is ideal for the expression of your cDNA of interest in mammalian tissue culture cells.

Library Preparation The cDNA used in making the library panels was synthesized from double purified mRNA using oligo(dT) primer. The cDNA was size fractionated on low melt agarose gels. The lanes were cut into multiple slices which were processed and ligated separately into the pCMV6-XL4 vector. The cDNA was directionally cloned into pCMV6-XL4 so that the CMV promoter will transcribe the cloned cDNAs. Each ligation was transformed and arrayed independently in the library panel. The libraries are arrayed so that larger sized cDNA clones are more highly represented in the arrayed panel. The libraries are arrayed so that the master plate contains plasmid miniprep DNA from all of the clones in the array. Each well on the master plate contains DNA from approximately 5000 clones, with a total of 500,000 clones in each library. Each individual well on the master plate corresponds to a single sub-plate. Each sub-plate well contains glycerol stocks of E. coli amplified from an original 50 clones.

Screening Procedure There are two basic ways to screen these libraries. The first is to use a specific pair of primers directed to your gene of interest to screen the library. The second method of screening is to vector primer PCR. The advantage of this second method is that you are able to identify which clones are larger. Which method you use depends on the quality of sequence information, location of the known sequence in the gene and size of the mRNA of interest. The vector primer is included with the each master plate.   Each master plate containes enough DNA for five complete library screens. The average insert size in all of our panels is between 3.0 and 4.5 kb. We recommend that you screen a panel which corresponds to a tissue where your gene of interest is expressed. The pattern of expression can be determined by Northern blot or PCR. To see a map of pCMV6 XL4

Human Library Panels

Library Panel Master Plate	Catalog Number
Brain (adult)	LAB-1001
Brain (fetal)	LLFB-1001
Heart	LHT-1001
Kidney	LKD-1001
Liver	LLI-1001
Lung	LLU-1001
Peripheral Blood Leukocytes (PBL)	LPBL-1001
Placenta	LLPC-1001
Skeletal Muscle	LMU-1001
Small Intestine	LSI-1001
Spleen	LLSP-1001
Human Testis	LTS-1001

Mouse Library Panels

Library Panel Master Plate	Catalog Number
Brain	MAB-1001
Embryo (12.5 day)	MEB-1001
Embryo (19 day)	MEA-1001
Liver	MLI-1001
Testis	MTS-1001
Thymus	MTM-1001

Rat Library Panels

Library Panel Master Plate	Catalog Number
Brain (adult)	RAB-1001

*Additional Panels coming soon

OriGene offers the Rapid-Screen™ "Longest-Clone" Screening Service for comprehensive screening of all human or mouse libraries listed above.

Sub-plates for the above libraries are designated by the library's prefix followed by the number (column) and letter (row) of the positive well(s) detected on the 96-well master plate, e.g. "RAB-5A"