Question

求助翻译！！！急！！

Qualifying the cDNA Library, continued

Expected
Digestion Patterns
Use the following guidelines to determine the size of the cDNA inserts. Refer to
page 70 for a sample electrophoresis.
• The pDONR™222 control will show a digestion pattern of 3 bands of the
following lengths:
2.5 kb
1.4 kb
790 bp
• Each cDNA entry clone should have a vector backbone band of 2.5 kb and
additional insert bands
• Make sure to digest enough plasmid DNA to be able to visualize smaller
insert bands (<300 bp)
• Make sure to run the gel long enough to distinguish bands representing
insert sizes of approximately 2.5 kb from the 2.5 kb vector backbone band

Determining
Average Insert
Size and %
Recombinants
1. Identify clones containing inserts using the guidelines outlined above.
2. For clones containing inserts, use the DNA ladder to estimate band sizes. If
there are multiple bands for a single cDNA entry clone, add all band sizes to
calculate the insert size. Do not include the 2.5 kb vector backbone band in
your calculations. Refer to page 70 for sample results.
3. Add together the insert sizes for all clones. Divide this number by the number
of clones containing inserts to calculate the average insert size for your cDNA
library.
4. Divide the number of clones containing inserts by the number of clones
analyzed to determine the percent recombinants.

What You Should
See
You should see an average insert size of ≥1.5 kb and at least 95% recombinants
for your cDNA library.
If the average insert size or percent recombinants of your library clones is
significantly lower, the cDNA going into the BP recombination reaction is either
of poor quality or is insufficient in quantity. For guidelines on isolating quality
mRNA, see page 10. To troubleshoot any of the cDNA synthesis steps, see
Troubleshooting, page 61.

The Next Step If you wish to sequence entry clones, proceed to Sequencing Entry Clones, next
page.
You may screen your cDNA library to identify a specific entry clone and use this
entry clone in an LR recombination reaction with a destination vector to generate
an expression clone. Refer to the Gateway® Technology manual to perform an LR
recombination reaction using a single entry clone.
Alternatively, you may transfer your cDNA library into a destination vector to
generate an expression library for functional analysis. For detailed guidelines,
refer to Performing the LR Library Transfer Reaction, page 57.
请帮忙翻译生物专业英语！！
谢谢！！！

Answer 1

合格DNA 图书馆, 继续

期望
消化样式
使用以下指南确定DNA 插入物的大小。参见
页70 为样品电泳法。
• pDONR™222 控制将显示3 条带的消化样式
随后而来的长度:
2.5 kb
1.4 kb
790 bp
• 各DNA 词条克隆应该有传染媒介中坚带2.5 kb 和
另外的插入物带
• 保证消化足够的质粒DNA 能形象化更小
插入带(

Answer 2