今天測序的結果返回了,是拼接過的,但是片段比我的目的片段大,是不是還含有載體序列?想通過DNAstar 的seqman去除載體序列,并拼接,用的是TransGen的pEASY-T1的載體,DNAstar中不包括,我自己添加了,但是設置不來啊,求幫助。pEASY-T1全長3928bp,說明書上描述如下:
LacZfragment: bases 1-547
M13 reverse priming site: bases 205-221
Multiple cloning site: bases 234-357
T7 promoter/priming site: bases 363-380
M13 Forward (-20) priming site: bases 388-403
f1 origin: bases 545-982
Kanamycin resistance ORF: bases 1316-2110
Ampicillin resistance ORF: bases 2128-2988
pUC origin: bases 3133-3806
以下是DNAstar里的幫助,我沒太看懂啊
Follow the steps below to modify a vector definition in the Vector Catalog:
1 From the Project Menu, select Vector Catalog.
2 Use the mouse to select the name of the vector you wish to edit.
3 Click Edit to open the Vector Definition window. Use the Tab key or the mouse to navigate between fields.
4 In the Vector field, edit the vector name.
5 Set a range for the vector sequence in the 5' and 3' coordinate boxes. You may input negative numbers. For instance, if the clone site were in position 75, you could enter -925 into the 5' box and 1075 into the 3' box to leave 1000 base pairs on either side without knowing the length of the vector. (我知道載體片段全長3928bp的話,那是應該設置-3928和3928嗎?)
6 In In the Clone Site box, enter the coordinates of the clone site and a range in the box. If you know the precise clone site, enter the position and use a range of 1. If you want a single definition for a number of sites in a polylinker, enter the midpoint position of the range of sites. If you use multiple sites in the same vector for different projects, you may define a separate vector entry for each clone site, provided you give each definition a different vector name. (載體說明書上寫的多克隆位點234-357,那這里應該怎么設置呢?)
7 Click OK to accept the modified definition.
求幫忙,糾結很久了,萬分感謝!
![DNASTAR分析測序結果]()
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