The following primers were used in

The following primers were used in 3’-RACE PCR to screen for defensins in M. menardi hemocytes together with the universal primer UPM: Spider def fwd 1, Spider def fwd 2, Spider def fwd 3, C.salei def fwd 1, C.salei def fwd 2, C.salei def fwd 3 and Polybetes def fwd. PCR with 1/100th of the synthesized cDNA of M. menardi. was performed with the following program: 5 min at 94°C, followed by 5 cycles of 94°C for 30 s, 51°C for 30 s, 72°C for 1 min, 5 cycles of 94°C for 30 s, 50°C for 30 s, 72°C for 1 min, 5 cycles of 94°C for 30 s, 49°C for 30 s, 72°C for 1 min, 5 cycles of 94°C for 30 s, 48°C for 30 s, 72°C for 1 min, 5 cycles of 94°C for 30 s, 47°C for 30 s, 72°C for 1 min, 5 cycles of 94°C for 30 s, 46°C for 30 s, 72°C for 1 min, and a final elongation at 72°C for 7 min . Candidate PCR products were run on a 0.7% agarose gel containing 0.5 ng ethidium bromide /ml. Single bands were cut out using a scalpel, and DNA was extracted from the gel using a MinElute® Gel Extraction Kit. Half of the extracted DNA was then used as template in another round of PCR, using the same primers and program as before. Products from this “nested” PCR were purified using the MinElute® PCR Purification Kit.

ผลลัพธ์ (อังกฤษ) 3:

The following primers were used in 3 '- RACE PCR to screen for defensins in M. Menardi hemocytes together with the universal. Primer UPM: Spider def FWD 1 Spider def, FWD 2 Spider def, FWD 3 C.salei def, FWD 1 C.salei def, FWD 2 C.salei def, FWD 3 and. Polybetes def FWD. PCR with 1 / 100th of the synthesized cDNA of M. Menardi. Was performed with the following program: 5 min. At 94 °, CFollowed by 5 cycles of 94 ° C for, 30 s 51 ° C for, 30 s 72 ° C for, 1 min 5 cycles of 94 ° C for, 30 s 50 ° C for, 30 s 72 ° C for 1 min 5 cycles,, Of 94 ° C for, 30 s 49 ° C for, 30 s 72 ° C for 1 min 5 cycles, of 94 ° C for, 30 s 48 ° C for, 30 s 72 ° C for 1 min 5 cycles, of 94 ° C. For, 30 s 47 ° C for, 30 s 72 ° C for, 1 min 5 cycles of 94 ° C for, 30 s 46 ° C for, 30 s 72 ° C for, 1 minAnd a final elongation at 72 ° C for 7 min. Candidate PCR products were run on a 0.7% agarose gel containing 0.5 ng ethidium. Bromide / ml. Single bands were cut out using a scalpel and DNA, was extracted from the gel using a MinElute apertium Gel Extraction. Kit. Half of the extracted DNA was then used as template in another round, of PCR using the same primers and program as. Before.Products from this "nested." PCR were purified using the MinElute apertium PCR Purification Kit.