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  1. Guide to Using Pooled Libraries

    Type
    Guide
    ...see if it should be amplified before you conduct your screen. If you need to amplify the library, please... Pooled Library Amplification Protocol for Addgene's generalized library amplification protocol. Read ...Overview of what pooled libraries are; how to amplify and use a pooled library; and what types of screens...: Pooled Library Guide Introduction Library Amplification Types of Screens Resources An Introduction to... 175,000 different gRNA-containing plasmids). Amplifying and Using the Library Once you receive your library...general steps to take in the screening process. Amplify the library (electroporation and maxiprep). If ...surviving cells are collected, their plasmids are PCR-amplified and sequenced using NGS to identify their cDNA...
  2. CRISPR Guide

    Type
    Guide
    ...experiments and must be amplified before use (Figure 8C), following the amplification protocol specified by... by the depositing lab. After amplification, you’ll need to use next-generation sequencing (NGS) to verify...verify that the composition of your amplified library matches that of the original library. Next, you’... copies of the aptamer improves stability and amplifies the signal for better imaging of genomic loci,...SHERLOCK (and SHERLOCKv2) allows for greater amplification and quantification and can be used on lateral... Innovative Genomics Institute, University of California, Berkeley. Plan Your CRISPR Experiment Resources... DNA PCR P olymerase C hain R eaction; Used to amplify a specific sequence of DNA pegRNA P rime E dit ...
  3. Plan Your Experiment

    Type
    Guide
    ...cell population. The region of interest is PCR amplified, PCR products are denatured-renatured, treated...restriction site. The region of interest is PCR amplified, digested with the appropriate restriction enzyme... agarose gel to identify DNA fragments. PCR amplification and gel electrophoresis (for HDR or NHEJ): For...insertions, the region of interest can be PCR amplified using primers that (A) flank the region of interest...determine whether the edit was successful. PCR amplification, subcloning and Sanger sequencing (for HDR or... sequence of targeted alleles. Involves PCR amplification of targeted region from DNA, subcloning into...plasmid, and screening individual clones. PCR amplification and next-generation sequencing (for HDR or NHEJ...
  4. Molecular Biology Reference

    Type
    Guide
    ... for the ability of the plasmid to be copied (amplified) by bacteria, which is an important characteristic...sequence used as an initiation point for PCR amplification or DNA sequencing of the plasmid. Primers can... using bacteria, which can uptake plasmids and amplify them during cell division. Because bacteria are... make proteins and other molecules essential for life. Scientists often “sequence DNA” to identify the...pieces, labeled with a short DNA barcode, and amplified. These DNA fragments are attached to a glass slide... other. These attached DNA templates are then amplified again producing ~1,000 copies of each template...
  5. Antibody Guide

    Type
    Guide
    ...Affinity Reagents Handling Visualization Signal Amplification Application Overview Quantification Methods ...present on the Fc region. This allows for the amplification of signals by having multiple secondary antibodies...on the antibody and conjugate chosen. Signal Amplification The amount of signal needed from a specific ...region of primary antibodies allows for signal amplification through the indirect detection method. In the...released from the complex and can be sequenced and amplified for analysis. Overview of application Process ...
  6. Cloning

    Type
    Guide
    ...discovery. With molecular cloning scientists can amplify and manipulate genes of interest and then insert...restriction sites to any piece of DNA during PCR amplification, allowing for it to then be digested and ligated...a single recombination reaction, drastically simplifying the process and reducing the amount of time required...). To do this your DNA fragment must first be amplified with specific Gateway attB1 and attB2 sites attached...
  7. Adenovirus Guide

    Type
    Guide
    ...replication-competent virus increases with each round of amplification. What is RCA? RCA stands for r eplication c ...crossover event increases with each round of amplification. Where can I find information about using the...
  8. Lentiviral Guide

    Type
    Guide
    ...interest using PCR. This is accomplished by PCR amplifying the insert sequence using primers that contain...
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