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  • 1. Cell Cloning By Serial Dilution In 96 Well Plates. Protocol ...

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Cell Cloning by Serial Dilution in 96 Well Plates Protocol Introduction John A. Ryan, Ph.D. Corning Incorporated Life Sciences 900 Chelmsford St Lowell, MA 01851 This technique is widely used for.

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You can calculate your cell concentration using the following formula: Total cells/ml = (Total cells counted x Dilution factor x 10,000 cells/ml)/ Number of squares counted. Total cells/ml = (325 cells x 2 x 10,000 cells/ml)/ 5 = 130 x 104 cells/ml. Total cells in sample = 130 x 104 cells/ml x 5 ml = 650 x 104 cells.

Limiting dilution cloning (LDC) We recommend performing limiting dilution by targeting 0.8 cells/well, which requires you to resuspend the transfected cells (post-counting) at a density of 8 cells/mL in complete growth medium, then transferring 100 μL of this to each well of a 96‑well plate.

Plate 200 µL of cell culture (i.e., 10,000–50,000 cells) into the wells of the sterile 96-well cell culture plate.

You have 3.348 x 10^6 cells. So if you need to seed 10000 cells per well, then for 100 wells you will need 10000 x 100 = 1 x 10^6 cells for a 96-well plate. If you seed cells at 100 ul per well, then you will need a total volume of 100 x 100 = 10000ul = 10 ml for 96-well plate.

You have 3.348 x 10^6 cells. So if you need to seed 10000 cells per well, then for 100 wells you will need 10000 x 100 = 1 x 10^6 cells for a 96-well plate. If you seed cells at 100 ul per well, then you will need a total volume of 100 x 100 = 10000ul = 10 ml for 96-well plate.

For any cell type you can use a minimum of 100ul to a maximum of 200ul culture medium per well in a 96-well plate.

Typically, 20,000 – 75,000 cells are seeded per well in Oris™ and Oris™ Pro 96-well plates, and 2,500 – 10,000 per well in Oris™ Pro 384-well plates.

To perform serial dilution, for example, 1 ml of the starting sample is added to 9 ml of Dilution Blank Tube 1. This is then followed by the same procedure, where 1 ml from Tube 1 is added to 9 ml of Tube 2, 1 ml from Tube 2 is added to 9 ml from Tube 3, and so on until the desired concentration is reached.

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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