Help me! info about gene cloning

Il Minn iminn at bioneer.kaist.ac.kr
Wed Feb 26 06:09:47 EST 1997


   I purified a protease, c.a. 80000Da, from Bacillus stearothermophilus
and determined the N-terminal sequences for gene clonong. Based on that 
sequences I designed the oligonucleotide primer for PCR. I used the total
genomic DNA as PCR template, but due to the high degeneracy of primer
I could not get correct PCR product. So my questions are as follows

1) Is there any method for increasing the specificity of PCR with highly
   degenerate primer?
2) Is there any method for making cDNA from Bacillus RNA?

3) Are there other methods for cloning a gene from Bacillus?

       I am very appreciate for your answers in advances

					Sincerely yours,
	                                Il Minn



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