Search found 5 matches

by coppcw08
Fri Nov 30, 2012 12:24 am
Forum: Molecular Biology
Topic: Cloning/Transformation question
Replies: 4
Views: 4339

Cloning/Transformation question

Hey all, I'm a noob at all this molecular stuff so I'm hoping you all can help a guy out. I ran a PCR today to amplify some genomic DNA that I will be cloning into a plasmid and transforming into some chemically competent E. coli cells. I was wondering what I should run as a negative control? I know...
by coppcw08
Tue Nov 20, 2012 6:59 pm
Forum: General Discussion
Topic: conversion
Replies: 1
Views: 1277

conversion

I have a primer that is 18.2 nmoles. I need to convert nanomoles to microliters.

I know the final answer is 182 microliters but that's because I know a shortcut on how to figure it out. Anyone know the "official" calculation/formula?

Thanks!
by coppcw08
Wed Nov 14, 2012 5:59 pm
Forum: Molecular Biology
Topic: TOPO PCR/Cloning
Replies: 1
Views: 1829

TOPO PCR/Cloning

I am going to be using TOPO cloning for the first time. I am using the Invitrogen p-ET-102 Directional cloning kit. My question is the following: Will I be using my entire recombinant protein? It says that I need a blunt-end PCR product but I'm not sure if that mean's I'll have my product chopped al...
by coppcw08
Mon Nov 05, 2012 12:27 am
Forum: Molecular Biology
Topic: Sequencing question/primer design
Replies: 4
Views: 5463

Re:

1) what he wants is primer for cloning, not for sequencing. Imagine you have sequence ATG GTC ATC GTC TGA GTC and first three amino acids belong to signal peptide (in reality it's much longer). In such case, you want your primer to start at GTC and go on further. By lipoprotein motif he probably me...
by coppcw08
Sun Nov 04, 2012 8:19 pm
Forum: Molecular Biology
Topic: Sequencing question/primer design
Replies: 4
Views: 5463

Sequencing question/primer design

My professor wants me to determine forward/reverse primers of a sequence (which I can do). However, he also wants me to determine sequencing primers (ones which will be in the middle of the sequence to provide complete gene coverage). There are 2 questions he is asking that confuses me. 1) Design a ...