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TypeBlog Post..., each flanked by a transcription start signal sequence (gene start) and a transcription termination/polyadenylation...luciferase or eGFP) flanked by the EBOV-specific gene start and gene end sequences. Thus, the minigenome contains...luciferase). The polymerase recognizes the gene start signal and initiates transcription of the reporter...
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Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)
TypeBlog Post...final vector structure you want to achieve before starting your experiments. Don’t dive directly into the... early stages of the process. Some questions to start with are: Is your sequence of interest a promoter...perform your MoClo assemblies in silico before you start doing your cloning. Most sequence analysis software... the line! Cash in on gene synthesis Before you start your cloning, it is advisable to remove unwanted... -
Viral Vectors 101: Preparing Pooled Libraries
TypeBlog Post...production steps before you can start transducing your virus. Let’s start there! Figure 1: ...services that can prepare the libraries for you. Starting from plasmids requires amplification, verification...disrupting plasmid representation. A good place to start is with a protocol for a library of similar size... -
Four Base Editing Reporters to Monitor and Enrich Editing in Real-time
TypeBlog Post...reporter proteins. Protein translation requires the start codon of AUG immediately downstream of a kozak sequence...generated a silent GFP construct, that contains an ACG start codon mutation, and integrated it into human and...As the initiation of protein translation at the start codon ATG is universal, the Dow lab successfully...vivo. As GO relies on the correction of a mutated start codon to initiate protein expression, it can be ... -
Troubleshooting and Optimizing a Western Blot
TypeBlog Post...forget to start it, the samples can actually migrate into the buffer — so it’s better to start over if ... runs to optimize or troubleshoot. Before you start… Mark your membrane! Membranes look the same from...optimize your antibody dilution further from this starting point. You can also consider using polyclonal ...than blots used to confirm overexpression. Before starting a western blot, think about how you’ll interpret... -
A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression
TypeBlog Post... longer than necessary before starting your experiment, or if you start an experiment before adequate ... or frt-STOP-frt (FSF) sequence prior to an ATG start codon to prevent transcription in cells where the...the serotype, except now you will already have a starting “optimal” injection volume and titer to begin ... -
Plasmids 101: Modular Cloning Applications and Kits
TypeBlog Post... uses of MoClo and the kits Addgene offers to kickstart these applications, no matter what your model ...isolation, then the MoClo Pichia Kit will get you started (Obst et al., 2017)! It is specifically curated...reporters (Engler et al., 2014). This kit is a great starter for your general plant cloning experiments. Addgene...with a large library of common parts to get you started quickly (Fonseca et al., 2019). Recently, the toolkit... -
How to Deposit Your Plasmids with Addgene
TypeBlog Post... need to create an account with Addgene before starting the process. After you have logged in to your ...Online Deposit" or "Download Deposit Spreadsheet". Start your plasmid deposit There are two options to begin...manuscript is published in a peer-reviewed journal. To start this process, simply enter your preprint title or... Learn the Benefits of Depositing with Addgene Start Your Deposit Browse Plasmids ... -
AAV Packaged on Request is Here!
TypeBlog Post...our new AAV Packaged on Request webpage to get started with your browsing and requesting. If you’re curious...easier to go from selection to viral vector. To start the AAV packaged on request process, simply navigate...placed, we’ll facilitate the necessary MTA, then start production and quality control. Our packaged on... -
Developing Transferable Skills During Science Training
TypeBlog Post... amazing students at Washington University who started the BALSA group, a nonprofit that provides short-term...stick to timelines Be a mentor in a formal program Start something (a journal, a science lunch club, a biotech...by Peggy Klaus Looking for a mentor or want to start a peer mentoring group? "What Makes a Good Mentor... -
CRISPR/Cas9 FAQs Answered!
TypeBlog Post...expression, is it necessary to add a G nucleotide to the start of my target sequence? A3: The human U6 promoter...promoter prefers a 'G' at the transcription start site to have high expression, so adding this G could help ...when they order the oligo. If your spacer sequence starts with a 'G', you naturally have one and do not need...that mutation be selected/screened? A13: Before starting your experiment, consider co-transfecting with... -
From our Table to Yours: An Inside Look at Lunch at Addgene
TypeBlog Post...Right from the start, when we founded Addgene nearly 10 years ago, lunch felt like a family meal. My ...We borrowed an idea from the software folks and started Hack Days. During Hack Days, teams at Addgene will...Executive Director, Joanne Kamens, responded by starting something we call “Random Lunches.” Once or twice... -
Bricking Science: Portraying Scientific Reality Through LEGO
TypeBlog Post... pathways just waiting to be discovered. As we started building, we thought – how can we make the connection...science video process One major challenge when we started was creating a LEGO stage that resembles a real...in a custom made, colorful laboratory. When we started, the hardest part of the process was to embed personality... -
Bringing Sustainable Practices to the Lab: Recycling
TypeBlog Post...States Postal Service picks up mail. NEB actually started this program in 1976! Millipore Sigma takes back...cost of regular waste services. Learn about how to start a glove recycling program at your institute! What...culture room for the wrappers and ship them back. Start a glove recycling program at your institute! Universities... -
Viral Vectors 101: Parts of the AAV Packaging Plasmid
TypeBlog Post...AAV. Where do you start? Plasmids of course! You just need three plasmids to start making the AAVs you...Alternative splicing and differing translational start sites generate the three VP proteins. The three ... -
Viral Vectors 101: Parts of the AAV Transfer Plasmid
TypeBlog Post...stop you. Where do you start? Turns out like all good experiments, making AAV starts with some plasmids. .... You just need three plasmids to start making AAV: the packaging plasmid which contains the structural... -
Mentoring for Scientists: I Have a Mentor, What Now?
TypeBlog Post...another person about their career path is a great start to a mentoring relationship, but if you don’t take...list available from this post. Use an activity to start a discussion. The Forced Choice Analysis is a fun...clarify and discuss values. The activity is just the start – it is the discussion after that will lead to learning... -
Networking Follow-up: I Met a Person. Now What?
TypeBlog Post...give rise to a starfish shape. You can ask me about the different arms of this starfish sometime, but... plug-in called Maps. Figure 1: See a starfish? This is actually a map of Joanne Kamen's LinkedIn... -
Virtual Science Conference Coming Up? Three Addgenies Share Their Experience
TypeBlog Post... questions,” says Zhang. “Usually the moderator starts by asking the speaker the highly upvoted question...chats between sessions or waiting for a session to start. “There was not much time to network,” Zhang says...discussing at tables and the event hasn't even started yet! @neurosymp will be interesting this year. ... -
5 Great Apps for Lab Life
TypeBlog Post... you. 2. Gibco Cell Culture by Gibco Whenever I start to learn a new technique, I try to look for a resource...for a mass spectrometry app. I digress. When I started learning proper cell culture technique, I tried...manual. Overall, it’s a great tool for someone starting cell culture work. 3. NEB Tools by New England...