| 1. | Then the pcr product was purified , ligated into pgem - t vector by ta cloning
筛选阳性克隆，测序，大量制备序列完全正确的质粒。 |
| 2. | The pcr product was inserted in e . coli clone vector pgem t - easy , and sequenced . a new subtype poifn - a was obtained
Pcr产物经纯化回收后，插入pgemt - easyvector ，经序列测定证实克隆了一新亚型猪干扰素。 |
| 3. | Pgem - t vector system , jm109 competent cells , reverse transcription kit and in vitro translation kit were purchased from promega company
Pgem一t载体，感受态细菌jm109 ，反转录试剂盒和体外翻译试剂盒均购自promega公司。 |
| 4. | Search the snps in wnk4 genomic sequences design the primers to get the unique pcr products of wnk4 gene which cover the full length genomic sequences
Dna的pgem一t载体为模板，用含有ha标签及酶切位点的上下游引物进行pcr扩增，酶切pedna3 |
| 5. | Put this fusion gene under the control of the promoter of ie1 gene , and then an eucaryotic cell expression plasmid vector pgem / ie1 " - gfp - actin resulted
将该融合基因置于ie1基因启动子的控制之下，构建成了真核表达质粒pgem ie1 - gfp - actin 。 |
| 6. | To make the pcr primer that have adequacy enzyme sit flowed the sequence , ligate with the pgem - t easy after pcr . make sure the sequence by sequencing
经组织培养后，以pcr和rt - pcr法检测再生烟草，共得到转synnhap基因烟草4株， pbi121空载体浸染的对照烟草再生苗5株。 |
| 7. | Using recombination techniques , the gene fragment ced - 9 , derived from pbs , was incorporated into pgem - 7zf ( + ) and formed a new vector pgem - ced9 , in order to get the right endonuclease sites
从质粒pbs中获取ced - 9基因片段，构建中间载体pgem - ced9以获得合适的酶切位点。 |
| 8. | Recombinant cloning vector of pgem - hc and pgem - ha were constructed by ligating the hc and ha genes into pgem t east vector and transformating into jm109 . 3
分别将目的基因hypoderminc和hypodermina与克隆载体( pgemteasy )连接并转化到宿主菌jm109中，构建了重组克隆载体pgem - hc和pgem - ha 。 |
| 9. | 2 . the special amplification products were ligated with the pgem - t easy vector and transferred into e . coli dh5 a via cohn method . after screening and identification , the positive clones were obtained
把扩增得到的特异片段连接到pgem - teasy载体上，用cohn法转化大肠杆菌dh5 ，经过筛选、鉴定，得到阳性克隆子。 |
| 10. | The amplified dna fragments were inserted into pgem - t easy vector and sequenced . the dna fragment sequencing results from the two subspecies were compared to detect whether there was any difference
将pcr扩增产物克隆到pgem - teasy载体，进行dna序列分析，并用生物信息学方法比较东方田鼠长江亚种与指名亚种之间该序列的差异。 |
