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Description
Organotypic balb/c mouse normal colon organoid culture KitProduct Specification Usage 1. Primary (1) the normal colonic tissue of balb c mice after sampling must be cleaned at 2 8 , several Petri dishes should be prepared, and primary culture buffer B pre cooled at 4 should be added for standby (double antibody and gentamicin should be added) (2) put the tissue into the Petri dish, wash it three times with primary culture buffer B, remove impurities, remove the intestinal mucosa of Balb c mouse colon tissue
Product Specification
| Usage |
1. Primary (1) the normal colonic tissue of balb/c mice after sampling must be cleaned at 2-8 ℃, several Petri dishes should be prepared, and primary culture buffer B pre cooled at 4 ℃ should be added for standby (double antibody and gentamicin should be added) (2) put the tissue into the Petri dish, wash it three times with primary culture buffer B, remove impurities, remove the intestinal mucosa of Balb/c mouse colon tissue with ophthalmic scissors or scalpel, and cut it into a tissue block with a volume of about 10-30mm3 (3) the tissues were digested with the digestive juice C of the primary normal colon tissues of balb/c mice, with shaking digestion at 4 ℃ for 20-25min or standing digestion for 40-50min (observe the digestion at any time during the digestion process) (4) take a small amount of liquid and observe under the microscope. More crypts are observed under the microscope. Add three times the volume of primary culture buffer B to stop digestion (5) filter with a 100um pore size screen, collect the filtrate, remove the supernatant after centrifugation at 300g for 5min, add primary culture buffer B and resuspend for centrifugation (6) Matrigel calculation: after step 5, observe the collected tissue volume, add 25 times the tissue volume of Matrigel (abs9495) to resuspend the plate (7) take a 24 well cell culture plate as an example, dispense 25ul of tissue Matrigel mixture per well for plating (operate at 4 ℃) (8) put the laid culture plate into a 37 ℃ incubator for 10-15min to form a gel, and add balb/c mouse normal colon organoid medium a (restore room temperature) for culture 2. Organoid subculture (1) pipette the culture medium, add 1-2ml of 4 ℃ organoid subculture buffer g to each well and place it for 2min (2) pipette the Matrigel gently, collect it in a 15ml centrifuge tube, and let it stand at 4 ℃ for 10min. (each 6-8 well is a group) (3) a: the number of organoids is insufficient or the volume is small: centrifuge for 5min to discard the supernatant, add an appropriate amount of organoid subculture buffer g to resuspend and transfer into a 1.5ml centrifuge tube, centrifuge for 5min at 300g to discard the liquid for step 4 b: When the number of organoids is large or the volume is large: centrifuge for 5min to discard the supernatant, add an appropriate amount of organoid passage digest D for digestion for 2-3min, add organoid passage culture buffer g to stop digestion, centrifuge for 5min to discard the mixture, add an appropriate amount of organoid passage culture buffer g to resuspend and transfer into a 1.5ml centrifuge tube, centrifuge for 5min at 300g to discard the liquid for step 4 (4) after organoid collection, add Matrigel to resuspend, lay 25ul Matrigel per well in a 24 well cell culture plate, place in the incubator for 10-15min, and add 500ul of balb/c mouse normal colon organoid culture medium A 3. Organoid cryopreservation (1) pipette the culture medium, add 1-2ml of 4 ℃ organoid subculture buffer g to each well and place it for 2min (2) pipette the Matrigel gently, collect it in a 15ml centrifuge tube, and let it stand at 4 ℃ for 10min. (each 6-8 well is a group) (3) centrifuge for 5min to discard the supernatant, add an appropriate amount of organoid subculture buffer g to resuspend, and centrifuge for 5min at 300g to discard the liquid (4) add an appropriate amount of organoid cryopreservation solution F, gently blow and resuspend. Take a 24 well cell culture plate as an example: the density is 2 wells and 1 tube is frozen, with a volume of 1.4ml per tube (5) mark the information, and after cooling down the program, transfer it to liquid nitrogen for long-term storage 4. Organoid resuscitation (1) take 10ml of organoid subculture buffer g and put it into a 15ml centrifuge tube (2) take out the frozen organoid cells from the liquid nitrogen tank and quickly put them into a 37 ℃ water bath pot for melting (3) during the water bath thawing process, it is necessary to gently shake the freezing tube to ensure that the frozen solution is completely thawed in 1-2min (4) quickly transfer the dissolved organoid cells to a 15ml centrifuge tube, gently blow 6-8 times with a pipette, centrifuge at 300g for 5min, and then remove the supernatant and collect the organoid cell pellet. Add an appropriate amount of organoid subculture buffer g to resuspend and transfer into a 1.5ml centrifuge tube. Centrifuge for 5min at 300g (5) resuspend Matrigel, lay 25ul Matrigel per well in a 24 well cell culture plate, place in an incubator for 10-15min to form a gel, and add 500ul of balb/c mouse normal colon organoid medium A. |
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| Synonym | Mouse(Balb/C) Colon Organoid Culture Kit | ||||||||||||||||
| Description | Composition:
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| Storage Temp. | 4 ℃, 3months; -20 ℃, 1 year, see the label for details |
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★★★★★ 5
great read
I enjoyed this book. As a person who enjoys history this book was a great read for me and the author gives an in depth vision on the trial.
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Reviewed in the United States on September 14, 2023
★★★★★ 3
Nat Turner was NOT happy
I think Mr Styron may have took some liberties with Nat Turner's "orientation" without the research or data to support his opinion.
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Reviewed in the United States on October 20, 2016
★★★★★ 5
A book for the ages...historically concise and horrific at once
William Styron, a gifted writer outdid himself with this book. His concise detail gives us an inside view on the depth of feeling employed by the subject of this book. The book while concise and tender pulls us in as these events take place. As Mailer delves deeper into the mind of Nat Turner, we cannot help but feel a sense of the suffering this man endured and this is what gives us perspective into this tragedy as it unfolds. A compelling masterpiece of literature.
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Reviewed in the United States on December 20, 2012
★★★★★ 5
Amazing Read and Crucial to Black Empowerment in America
Format: Paperback
This book should be in every black home in America, it shows in clear and evident words the flaws in America towards African Americans even into the modern day, as well as the reasons for the various obstacles to progress and change for the Black Community. An all around well-rounded read for any Black soul that is a must have to understand the reality they will endure and how they can achieve the reality they deserve.
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Reviewed in the United States on January 12, 2025
★★★★★ 5
Good gift but particular gift for this difficult person to shop for
Format: Paperback
Got this as a gift for a friend and he was hard to shop for even after asking his wife. He's not a handyman, or a tech savvy man. He cooks, and is a well read man with interests in politics and psychology, he's not at all a gamer either. He doesnt DO much but work, it's not like he works out, golf's or anything like that... And they have a Brady bunch family and all she gave me to go on was he likes malcol x. And this author is more Malcom x than mlk and aggressive in views (when being compared to MLK) but is also very intertwined with their philosophies as he was IN it... According to the person I gave the gift to. Basically he loved it! So
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Reviewed in the United States on January 14, 2019
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