Construction of human VL gene library [Equipment and reagents] ◠PCR reagents and equipment ◠cFv gene library single-stranded template DNA, prepared from natural scFv library in pHENl (10ng / u1) ◠Gelleclean kit (Qbiogene) ◠Wtzard PCR purification kit (ProlneSa) ◠RJHl / 2Xho primer: 5-GGC ACC CTG GTC ACC GTC TCG AGT GGT GGA-3 ◠RJH3Xho primer: 5-GGG ACA ATG GTC ACC GTC TCG AGT GGT GGA-3 ◠RJH4 / 5Xho primer: 5-GGA ACC CTG GTC ACC GTC TCG AGT GGT GGA-3 ◠RJH6Xho primer: 5-GGG ACC ACG GTC ACC GTC TCG AGT GGT GGA-3 ◠FdSEQl primer ◠pHENl-Vλ3 vector DNA (according to MTA) [method] 1. Prepare 4 independent 50ul PCR reaction mixtures, including: ◠Water, 35.5ul ◠20XdNTP (each 5mmol / L), 2.5ul ◠10XVent polymerase buffer, 5.0ul ◠FdSEQl primer (10 pmol/u1), 2.5ul ◠Reverse primer (10 pmol/u1), 2.5ul ◠Single-stranded scFv template DNA (10ng), 1.0ul ◠Vent DNA polymerase (2 units), 1.0ul 2. Heat the reaction mixture to 94 ° C for 5 minutes in a thermal cycler with a heated lid. 3. Amplify the VL gene by cycling a total of 25 times at 94 ° C for 30 seconds, 42 ° C for 30 seconds, and 72 ° C for 1 minute. 4. Use the Wizard PCR purification kit to purify the PCR product. 5. Digest the PCR product with XhoI and NotI; but in addition to using XhoI instead of NcoI, NEB2 buffer and BSA are required, in accordance with the manufacturer's requirements. 6. Digest pHENl-VL3 vector DNA with XhoI and NotI; but do not replace NcoI with XhoI. 7. Connect the digested PCR product and digested vector, and electrotransform them into E. coli TG1 cells to construct 4 VL gene phage libraries. The library capacity was measured and the bacterial library material was stored at -70 ° C. 8. Inoculate 100ml 2X TY medium containing 100ug / ml ampicillin and 1% glucose with bacteria containing glycerol library storage material to prepare each VL gene library DNA. The inoculation amount should be large enough to ensure that the number of inoculated bacteria is at least less than the library The capacity is 5 times larger. After incubating overnight, follow the standard DNA plasmid preparation method. For subsequent library digestion, DNA from different libraries can be combined. Solid Wood Furniture,Solid Wood Office Furniture,Solid Wood Nightstand,Solid Wood Bedroom Furniture Hebei Rukai Furniture Co. , Ltd. , https://www.rukaifurniture.com