TAIL PCR (CAT#: STEM-MB-0194-WXH)

TAIL PCR utilizes three nested primers in consecutive reactions together with an arbitrary degenerate primer having a low melting temperature so that relative amplification frequencies of specific and non-specific products can be thermally controlled.

• Amplifying insert end segments from P1, BAC and YAC clones.
• Direct Sequencing.
• Recovery single-copy sequences from highly complex genome.
• Rapid isolation of promoter sequences.

TAIL-PCR is generally divided into 3 reactions.
1. The first PCR reaction consists of 5 high specificity reactions, 1 low specificity reaction, 10 lower specificity reactions and 12 thermally asymmetric supercycles. Three types of products were obtained at different concentrations after the above reactions: specific product (type I) and non-specific product (type II and III).
2. In the second reaction, the product of the first stage reaction was diluted 1000 times as a template, and the specific product was selectively amplified while the non-specific product was suppressed to a very low level by 10 thermally asymmetric supercycles.
3. The third reaction, in which the product of the second reaction is diluted 1000 times as template, is generally set up as a normal PCR reaction or a thermally asymmetric supercycle.

Three nested specific primers (special prime, sp1, sp2, sp3, about 20 bp) were combined with one short (14 bp) random degenerate prime (AD) with low Tm value, respectively.