What is a digest?
A digest is a compilation of case annotations, arranged alphabetically by subject. Case annotations are short descriptions of a single aspect of a case. Most digests are published by West/Thomson Reuters and use the West Topic and Key Number System.
What is meant by single digestion of DNA?
Hence, this process is called single digestion. Generally, plasmids used in recombinant DNA technology only contains a single restriction site for a particular restriction enzyme. Therefore, single digestion makes the circular plasmid linear, yielding a single DNA fragment.
What are single-digested plasmids?
Single-digested plasmids are a type of plasmids digested only with a single restriction enzyme. Hence, this process is called single digestion. Generally, plasmids used in recombinant DNA technology only contains a single restriction site for a particular restriction enzyme.
What are restriction digests and how do they work?
Given the variety of these enzymes and the unique sites they recognize, restriction digests have become the most widely used method scientists employ to selectively move a specific piece of DNA from one plasmid to another.
What is a double digestion?
Digesting a DNA substrate with two restriction endonucleases simultaneously (double digestion) is a common timesaving procedure. Selecting the best NEBuffer to provide reaction conditions that optimize enzyme activity as well as avoid star activity associated with some enzymes is an important consideration.
What is a digest in genetics?
Restriction Digestion is the process of cutting DNA molecules into smaller pieces with special enzymes called Restriction Endonucleases (sometimes just called Restriction Enzymes or RE's).
Why do we do double digest?
So, if the two RE's are compatible bufferwise you might do a double digest in one go, but depending on the compatibility of your two RE's of choice, you might need to do it in two phases: stop the reaction of the first digest (heat-shock), purify the DNA-digest, then do the second digest...
What is a digest in gel electrophoresis?
The goal of a diagnostic digest is to cut your plasmid into specific sized pieces and analyze the resulting fragments by gel electrophoresis. The pattern of the fragments on the gel can indicate if the plasmid contains the expected size insert.
What is a plasmid digest?
Purified plasmid DNA is digested with 1 or more restriction enzymes (REs) selected to give a distinct DNA band pattern that is easily resolved by electrophoresis.
Why do a restriction digest?
Restriction digestion is usually used to prepare a DNA fragment for subsequence molecular cloning, as the procedure allows fragments of DNA to be pieced together like building blocks via ligation.
How long can you leave a restriction digest?
If you are proceeding with downstream experiment like ligation and for cloning it is advisable to digest maximum of 4 hours or over night with less unit of enzymes.
What is partial digestion?
Biology Glossary search by EverythingBio.com. A restriction digest that has not been allowed to go to completion and thus contains pieces of DNA with some restriction endonuclease sites that have not yet been cleaved.
What happens if you use too much restriction enzyme?
Incomplete digestion is a frequently encountered issue when using restriction endonucleases. Incomplete digestion may occur when too much or too little enzyme is used. The presence of contaminants in the DNA sample can inhibit the enzymes, also resulting in incomplete digestion.
What is digestion in PCR?
The most convenient option for digestion of PCR-ampli- fied DNA is the addition of a restriction enzyme directly to the reaction tube after completion of PCR. The majority of restriction enzymes are active in PCR buffers. However, digestion of PCR products in the amplification mixture is often inefficient.
Why is SDS used in gel electrophoresis?
In SDS-PAGE, the use of sodium dodecyl sulfate (SDS, also known as sodium lauryl sulfate) and polyacrylamide gel largely eliminates the influence of the structure and charge, and proteins are separated solely based on polypeptide chain length.
Why do we digest DNA?
The resulting digested DNA is very often selectively amplified using polymerase chain reaction (PCR), making it more suitable for analytical techniques such as agarose gel electrophoresis, and chromatography. It is used in genetic fingerprinting, plasmid subcloning, and RFLP analysis.
What does digest mean in science?
English Language Learners Definition of digest (Entry 2 of 2) : to change (food that you have eaten) by a biological process into simpler forms that can be used by the body. : to think over and try to understand (news, information, etc.) See the full definition for digest in the English Language Learners Dictionary.
What does digest mean in medical terms?
Medical Definition of digest (Entry 2 of 2) 1 : to convert (food) into absorbable form. 2 a : to soften, decompose, or break down by heat and moisture or chemicals. b : to extract soluble ingredients from by warming with a liquid. intransitive verb.
What does "digestion" mean for kids?
Kids Definition of digest (Entry 2 of 2) 1 : to change or become changed into simpler forms that can be used by the body digest a meal My dinner is still digesting. 2 : to think over and try to understand That's a lot of information to digest.
What does "digerere" mean in Latin?
Middle English, systematic arrangement of laws, from Latin digesta, from neuter plural of digestus, past participle of digerere to arrange, distribute, digest, from dis- + gerere to carry. Verb. Middle English, from Latin digestus. Keep scrolling for more.
What is digest case?
A digest is a compilation of case annotations, arranged alphabetically by subject. Case annotations are short descriptions of a single aspect of a case. Most digests are published by West/Thomson Reuters and use the West Topic and Key Number System. This system allows the researcher to research similar cases in any jurisdiction ...
What is the West's American Digest System?
West's American Digest System (11th Decennial Digest, South Wing, KF 141 1996) covers cases from all fifty states and all federal courts, including the U.S. Supreme Court. This is a very useful source when you need to locate cases from multiple jurisdictions that deal with the same issue.
Why is the topic and key number system used in West's online system?
Because the topic and key number system from the print digests and reporters is also used in West's online system, knowing how to locate case law in print, and understanding how the information is organized, can lead to better online research results. Sometimes when online research is unsuccessful browsing through a print digest will help ...
What is the difference between a hash and a digest?
The basic difference between a hash function and digest is that digest is the value obtained from a hash function. A hash function is any function that can be used to map data of arbitrary size to data of fixed size. The values returned by a hash function are called hash values, hash codes, digests, or simply hashes.
What is digest in hash?
The digest is the output of the hash function. For example, sha256 has a digest of 256 bits, i.e. its digest has a length of 32 bytes. That's it really. The basic difference between a hash function and digest is that digest is the value obtained from a hash function.
What is diagnostic digest?
The goal of a diagnostic digest is to cut your plasmid into specific sized pieces and analyze the resulting fragments by gel electrophoresis. The pattern of the fragments on the gel can indicate if the plasmid contains the expected size insert.
What is restriction digest?
Given the variety of these enzymes and the unique sites they recognize, restriction digests have become the most widely used method scientists employ to selectively move a specific piece of DNA from one plasmid to another.
Most recent answer
I've seen the phenomenon that Donna describes too. Not to mention white colonies without an insert.
Popular Answers (1)
You have 50% chance to have your insert in the correct orientation. So why are you complicating the task just do your cloning and screen for the right orientation using PCR and pair of Primers one on the vector and one in the right direction of your insert. All the positive clones will give a band with the expected size
All Answers (13)
You have 50% chance to have your insert in the correct orientation. So why are you complicating the task just do your cloning and screen for the right orientation using PCR and pair of Primers one on the vector and one in the right direction of your insert. All the positive clones will give a band with the expected size
Similar questions and discussions
How much DNA template (genomic or plasmid DNA) is used for a general PCR?
What is restriction enzyme digestion?
Restriction enzyme digestion takes advantage of naturally occurring enzymes that cleave DNA at specific sequences. There are hundreds of different restriction enzymes, allowing scientists to target a wide variety of recognition sequences. For a list of many commonly used restriction enzymes, visit NEB .
How long to digest DNA for cloning?
For diagnostic digests, 1-2 hours is often sufficient. For digests with >1 µg of DNA used for cloning, it is recommended that you digest for at least 4 hours. *Pro-Tip* If you will be using the digested DNA for another application (such as a digestion with another enzyme in a different buffer), but will not be gel purifying it, ...
