m******t
发帖数: 109 | 1 i plan to construct a plasmid with 3 amino acid mutations. anyone with
experience? please help. i appreciate it very much..... |
a*****n
发帖数: 2835 | 2 check stratagene website
they have a program for designing primers. I did a lot mutagenesis using
their program and system.
【在 m******t 的大作中提到】
: i plan to construct a plasmid with 3 amino acid mutations. anyone with
: experience? please help. i appreciate it very much.....
|
m******t
发帖数: 109 | |
a***a
发帖数: 40617 | 4 quikchange
【在 m******t 的大作中提到】
: thx very much.
|
x*p
发帖数: 6 | 5 顺路问一下，order的primer的时候一定要PAGE purification吗？ 谢谢 |
k*******s
发帖数: 214 | 6 Contact me. We can finish multiple mutation in once cloning. Thanks |
m******t
发帖数: 109 | 7 2 questions: i was provided 2 pairs of primers for 3 mutations by the
stratagene program. 1' i should use the MULTI kit cause there are several
kits. 2' should i do 1 or 2 PCR? thx
【在 m******t 的大作中提到】
: i plan to construct a plasmid with 3 amino acid mutations. anyone with
: experience? please help. i appreciate it very much.....
|
e****s
发帖数: 1125 | 8 说说看怎么做的？
Overlapping PCR?还是什么Kit?
【在 k*******s 的大作中提到】
: Contact me. We can finish multiple mutation in once cloning. Thanks
|
m******t
发帖数: 109 | 9 i am going to do mutations. kit and primers are ready. however, still have
questions. please help.
1. i got 2 PAIRS of primers (sense and anti-sense for each pair). when i
read the manual, i found that for the control, they used 3 primers for 3
mutations, one single primer for one mutation. so i am confused now. should
i use pairs of primers or just sense or anti-sense primer?
2 may i use common SOC medium or i have to use NZY+ both for XL10 competent
cells?
any other suggestions are greatly a
【在 m******t 的大作中提到】
: i plan to construct a plasmid with 3 amino acid mutations. anyone with
: experience? please help. i appreciate it very much.....
|
