input SELEX libary for AntHox6a
SELEX library with a 16-mer randomized region whose full sequence is as follows: GTTCAGAGTTCTACAGTC-CGACCTAA-16N-TTAGG-ACTCGGACCTGGACTAGG The libraries were double strand DNA annealed and extended with Klenow polymerase. The SELEX annealing primers are as follows: SELEX-L - GTTCAGAGTTCTACAGTCCGA SELEX-R CCTAGTCCAGGTCCGAGT To be specific, the initial mixture was prepared in a DNA LoBind Safe lock tube (Eppendorf 022431021) by combining 10L of 10x STE buffer (100mM Tris pH8.0, 10mM EDTA pH8.0, 1M NaCl), 10L of 100M SELEX library oligo, 20L of 100M annealing primer (sequence), and 60L of water. The tube was then placed in a 1-liter beaker containing 800mL of water and boiled for 10 minutes. After boiling, the tube was left in the hot water until the water cooled to room temperature naturally. Next, 100L of the annealed DNA from the first step was taken and mixed with 25L of 10x Klenow buffer, 20L of 10mM dNTP, 80L of water, and 20L of Klenow fragment (NEB M0210L). The mixture was incubated at room temperature for 30 minutes. To stop the reaction, 10L of 0.5M EDTA at pH 8.0 was added. Following this, 25L of 3M sodium acetate at pH 5.2 was added, and the DNA was purified using a Qiagen column (Qiagen 28106) according to the regular PCR purification protocol.