Hi

I am a student and clarify the following.

1. Is that possible to obtain complete coding sequence of cdna of our gene of interest commercially?

2.Can a pcr product of particular gene after the gel extraction be used as template for cdna synthesis directly ? (The cdna is intended to be used for transfection after inserting into expression vector).

Please reply, even if i don't sound sensible.

many cdna's can be obtained commercially, could be pcr'ed , inserted into a vector and transfected into a cell.

Here are some online resources for cDNA clones:

OriGene - cDNA Cloning and Selectable Vectors

OriGene - TruClone Human Full-Length cDNA Clones - Application Guide

U of Missouri-Rolla cDNA Resource Center from Biochain




Here are some links to a cDNA transfection methods

Introduction to cDNAProtocol: cDNA Transfection using FuGENE 6 (Boehringer Mannheim)


Cells-to-cDNA II Applications

Ambion's Cells-to-cDNA II - Instruction Manual

Ambion's Technote 9(3) - Synthesize cDNA Directly From Limiting Samples

An easy way to obtain the cDNA for your gene of interest is to search for your gene using the "core nucleotide" feature of NCBI - http://www.ncbi.nlm.nih.gov/sites/entrez

For example. I will look for gene "CACNG2" from human

Look down the results list for a Human hit and on the right hand side some have the link to "order cDNA clone". [CLICK TO SEE A BIGGER VERSION]

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I have highlighted a good hit

Click the Order cDNA clone link and you will have a choice of sources like Geneservice Ltd, Open Biosystems, RZPD German Resource Center for Genome Research, and ATCC. Each vendor will have the cDNA in a variety of vectors so choose one which is going to mean the least subcloning for you. If you luck out they sometimes have them in pCMV based vectors so they are ready for mammalian expression.

Typically clones cost between $75 and $100 depending on the vendor and other factors