連接擴增反應 的英文怎麼說
中文拼音 [liánjiēkuòzēngfǎnyīng]
連接擴增反應
英文
ligation amplification reaction- 連 : Ⅰ動詞1 （連接） link; join; connect 2 （連累） involve （in trouble）; implicate 3 [方言] （縫） ...
- 接 : Ⅰ動詞1 （靠近;接觸） come into contact with; come close to 2 （連接; 使連接） connect; join; put ...
- 擴 : 動詞（擴大） expand; enlarge; extend
- 反 : Ⅰ名詞1 （方向相背） reverse side 2 （造反） rebellion 3 （指反革命、反動派） counterrevolutionari...
- 應 : 應動詞1 （回答） answer; respond to; echo 2 （滿足要求） comply with; grant 3 （順應; 適應） suit...
- 連接 : connect; fit together; link; marry; mate; joint; association trail; linkage; concatenate; concate...
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After electrophorised on 1 % agarose gel, the pcr production was purified with agarose gel dna extraction kit. the segment was ligated with vector pmd18 - t and then was tranformed into the competent cell of dh5 a. a construction mstnd - pmd18t was generated by inserting the sequence of 254bp into pmd18 - t vector and selecting the sense clones. positive clone was identified by three ways : endonuclease digestion, pcr and sequencing. the result showed that the cloned sequence coincides with the designed sequence. this construction was digested with nco i and xho i and ligated the pet28a ( + ) vector digested with the same enzymes using dna ligation kit. the production of ligation reaction was transformed into the competent cell of bl21 ( de3 ). after 12 - 16 hours of culture, several colnes appeared on the plate. some positive clones were selected to extract their plasmid. these plasmids were digested by nco i and xho i and indentified by pcr. a contraction, mstnd - pet28a was generated. the result showed that the cloned sequence coincides with the designed sequence
F _ 1長38bp ， r _ 1長36bp ，其它片段均40bp長， f _ 1和r _ 1片段兩端分別加上限制性內切酶nco和xho的識別位點序列。用成對單鏈片段進行延伸反應，然後用其他單鏈片段作為引物，進行pcr擴增，用dna快速純化回收試劑盒回收所得254bppcr產物，與pmd18 - t載體連接、轉化dh _ 5 。受體菌感受態細胞，利用藍白斑遺傳學篩選法篩選陽性克隆，提取其質粒，採用nco和xho雙酶切鑒定，獲得了254bp的片段；用pmd18 - t載體上的特異引物rv - m和m13 - 47進行pcr鑒定，獲得300bp的片段。
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