《Table 1 Primer pairs used to amplify and detection Foxn1 gene》

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《Evolution and Expression Patterns of Forkhead Box N1 in Pig》

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Based on the reference sequence of the pig Foxn1gene（Gen Bank Acc.No.:NC_010460）in NCBI，which was derived from automated computational analysis，10 primer pairs were designed to clone its DNA sequence.One primer pair（CDSF and CDSR）was designed to clone the entire CDS.The sequence and location of the primers are shown in Table 1.PCR reaction system（the total reaction volume:25μL）included 1μL of genomic DNA（75 to 150 ng），2.5μL1×Taq reaction buffer with Mg Cl2，2μL of d NTPs（0.2 mmol·L-1），1μL each primer（0.2μmol·L-1），0.2μL of r Taq DNA polymerase（1 U）and 17.3μL R N a s e-f r e e H2O.P C R w a s c o n d u c t e d b y p r edenaturing at 95℃for 5 min；followed by 30 cycles of 95℃for 1 min，55-60℃for 1.5 min，and 72℃for1 min；and a final extension at 72℃for 10 min.All the PCR products were detected by 1%agarose electrophoresis and purified using a gel extraction kit.The ligation reaction system contained 0.5μL of p MD18-T vector，5μL of the purified PCR products and 4.5μL of the ligation buffer，which was then incubated at 4℃for at least 12 h.All the reagents were purchased from Ta Ka Ra.Cloning products were sequenced by Huada Biotechnology Co.，Ltd.