今天测序的结果返回了,是拼接过的,但是片段比我的目的片段大,是不是还含有载体序列?想通过DNAstar 的seqman去除载体序列,并拼接,用的是TransGen的pEASY-T1的载体,DNAstar中不包括,我自己添加了,但是设置不来啊,求帮助。pEASY-T1全长3928bp,说明书上描述如下:
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里的帮助,我没太看懂啊
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. (我知道载体片段全长3928bp的话,那是应该设置-3928和3928吗?)
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. (载体说明书上写的多克隆位点234-357,那这里应该怎么设置呢?)
7 Click OK to accept the modified definition.
求帮忙,纠结很久了,万分感谢!
![DNASTAR分析测序结果]()
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