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O Describe what a plasmid is. o Describe the function of the three essential features of all cloning plasmids. o Explain how electrophoresis of DNA works. o Explain how electrophoresis can be used to determine the size of a fragment of DNA. I. Objectives: Isolate a plasmid containing a cloned soybean gene. Use restriction enzymes to release the soybean gene from the plasmid. Use gel electrophoresis to determine the size of the soybean gene. II. Background: Plasmids Bacteria have a s.

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Plasmid extraction uses size-based separation, while genomic DNA extraction involves lysis and purification via ethanol precipitation.

DNA on Filter Paper To recover the plasmid, use a clean razor blade to cut out one of the circles containing your DNA. Immerse the circle in 30 µL of TE and pipette to mix. After waiting for at least 10 minutes, use 2 µL to transform competent bacteria.

EtBr can affect the conformation of your plasmid, obscuring the perspective on your sample. EtBr is typically used at concentrations of 0.2-0.5 ug/mL, and while increasing concentrations significantly does not affect bands for smaller plasmids, larger plasmids can exhibit variations in number of bands and positions.

Incubate the culture overnight at 37°C with vigorous shaking. Pour 1.5 ml of the culture into a microfuge tube. Centrifuge at maximum speed for 30 seconds at 4°C in a microfuge. Store the unused portion of the original culture at 4°C.

The basic steps of plamid isolation are disruption of the cellular structure to create a lysate, separation of the plasmid from the chromosomal DNA, cell debris and other insoluble material.

Key points: Gel electrophoresis is a technique used to separate DNA fragments ing to their size. DNA samples are loaded into wells (indentations) at one end of a gel, and an electric current is applied to pull them through the gel. DNA fragments are negatively charged, so they move towards the positive electrode.

10.3: Plasmids are easily isolated from bacterial cells. Plasmid isolation takes advantage of the unique structural properties of plasmids. Plasmids are small, supercoiled circular pieces of DNA. Unlike the much larger bacterial chromosome (which is also circular), plasmids are quite resistant to permanent denaturation ...

A lysis solution containing sodium hydroxide (NaOH) and a detergent (such as SDS) is added to the resuspended bacteria. The mixture is gently mixed and incubated for a short period. This step results in the breakdown of the cell wall and membrane, releasing the cell's contents, including plasmid DNA, into the solution.

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Form Packages
Adoption
Bankruptcy
Contractors
Divorce
Home Sales
Employment
Identity Theft
Incorporation
Landlord Tenant
Living Trust
Name Change
Personal Planning
Small Business
Wills & Estates
Packages A-Z
Form Categories
Affidavits
Bankruptcy
Bill of Sale
Corporate - LLC
Divorce
Employment
Identity Theft
Internet Technology
Landlord Tenant
Living Wills
Name Change
Power of Attorney
Real Estate
Small Estates
Wills
All Forms
Forms A-Z
Form Library
Customer Service
Terms of Service
DMCA Policy
About Us
Blog
Affiliates
Contact Us
Privacy Notice
Delete My Account
Site Map
All Forms
Search all Forms
Industries
Forms in Spanish
Localized Forms
Legal Guides
Real Estate Handbook
All Guides
Prepared for You
Notarize
Incorporation services
Our Customers
For Consumers
For Small Business
For Attorneys
Our Sites
US Legal Forms
USLegal
FormsPass
pdfFiller
signNow
airSlate workflows
DocHub
Instapage
Social Media
Call us now toll free:
1-877-389-0141
As seen in:
  • USA Today logo picture
  • CBC News logo picture
  • LA Times logo picture
  • The Washington Post logo picture
  • AP logo picture
  • Forbes logo picture
© Copyright 1997-2025
airSlate Legal Forms, Inc.
3720 Flowood Dr, Flowood, Mississippi 39232