Whole gene cloning - (Sep/07/2005 )
Dear, all
I want to clone whole gene in fungi. Now, I have partial of this gene. (around 200 bp). The whole gene is around 1-2 kb. Please suggest me which method is good for getting the whole gene and please give deeply detail of the method.
Thank you for your advise
I want to clone whole gene in fungi. Now, I have partial of this gene. (around 200 bp). The whole gene is around 1-2 kb. Please suggest me which method is good for getting the whole gene and please give deeply detail of the method.
Thank you for your advise
Does RACE-PCR work with fungi? It should...I'd select an RLM method for RACE. Sequence the 200 bp and design primers for amplification using RACE. Then get a kit for it. It should cost about $700, I believe.
-Matt
This is project depending on what you have to start with and what means more to you - time or money? What's the species? Has it been sequenced? If so peruse online data sources with your 200bp sequence (NCBI, TIGRE, etc).
Query your 200bp against published sources to find the coding or genomic sequence. Use this to design PCR primers and amplify your gene from genomic DNA for full sequence or cDNA for just the cds.
You can probably get away with using sequence from a species in the same genus as well if you species has not been sequenced.