Looking for a RACE PCR KIT that enables Products amplification and sequencing without the cloning step .Do you know if such a method or kit is available or can you recommend one?
I found the: 5’/3’ RACE Kit, 2nd Generation from Roche that allows RACE PCR products sequencing without cloning. Has anyone tried it? Any thoughts?
Hi Nina
Most RACE kits will allow amplification of the products without need for cloning right in the begining.....but at certain stage you need to clone the products and sequence them to see..if you got what you want..
bye
Normally, product amplified by RACE PCR kit need to be cloned into specific vector like pGEM-T easy vector before the purified recombinant plasmid containing the product of interest for sequencing. The RACE products can be cloned into an appropriate vector for subsequent characterization studies.
If you get a clean single band when performing ordinary PCR, sequencing the PCR product can save time over cloning. However,if you do not have a clean single band. you are doing RACE with specificity only at one end, which reduces the likelihood of getting a specific product.
Otherwise, products of the RACE reaction from that kit can be directly sequenced without cloning.
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