What is Lambda HindIII DNA marker?
Thermo Scientific Lambda DNA/HindIII Marker is recommended for sizing of linear double-stranded large DNA fragments in agarose gels. Lambda DNA is digested to completion with the appropriate Thermo Scientific restriction enzyme(s) and purified and dissolved in storage buffer.
How many bands are there in HindIII?
The Lambda DNA is completely digested with HindIII to yield bands ranging from 0.125 kb to 23 kb, suitable for use as molecular weight standards for agarose gels. The marker is composed to 8 purified individual DNA fragments (in base pairs): 23130*, 9416, 6557, 4361*, 2322, 2027, 564 and 125.
How many times does HindIII cut lambda DNA?
Description: The HindIII digest of lambda DNA (cI857ind 1 Sam 7) yields 8 fragments suitable for use as molecular weight standards for agarose gel electrophoresis (1).
What is lambda DNA?
Lambda DNA, a linear, double-stranded phage DNA containing 12 bp single-stranded complementary 5′-ends, is derived from an Escherichia coli bacteriophage (Bacteriophage lambda cI857 Sam7). Lambda DNA can also be used as a substrate in restriction enzyme activity assays.
What is the restriction site for HindIII?
Thermo Scientific HindIII restriction enzyme recognizes A^AGCTT sites and cuts best at 37°C in R buffer. See Reaction Conditions for Restriction Enzymes for a table of enzyme activity, conditions for double digestion, and heat inactivation for this and other restriction enzymes.
What is Lambda HindIII ladder?
The λ HindIII DNA Ladder is prepared by restriction digestion of phage Lambda DNA to completion with HindIII, followed by heat inactivation of the enzyme. The resulting 7 fragments range from 564 base pairs to 23.13 kilobases in size. The λ Hind DNA Ladder is supplied ready to use in loading buffer containing dye.
How many bands are visible in the digested lambda DNA?
Lambda phage-DNA has 5 EcoRI restriction sites so after it should produces 6 bands but I can see only 4 out of 6 possible bands.
What is Lambda ladder?
The lambda ladder consists of suc- cessively larger concatemers of λ cl857 Sam7. These DNA size standards may be used as size markers for CHEF, FIGE, and other pulsed field electrophoresis gels (Figure 1). Quantity. Five agarose inserts (25–40 lanes).
Why is lambda DNA important?
It binds DNA in the helix-turn-helix binding motif. It regulates the transcription of the cI protein and the Cro protein. The life cycle of lambda phages is controlled by cI and Cro proteins.
How many ecor1 sites are there in lambda DNA?
5 recognition sites
The Lambda DNA used in this experiment is isolated as a linear molecule from the E. coli bacteriophage lambda. It contains approximately 49,000 base pairs and has 5 recognition sites for Eco RI, and 7 for Hind III.
What does the III in HindIII stand for?
(iii) The Roman numerats denoted the sequence in which the restriction endonucleas e enzyme from that particular genus, species and strain of bacteria have been isolated iii i.e., third retriction endonuclease to be isolated from this species. Question Details till 21/01/2022.
How do you separate DNA fragments from lambda phage?
In this experiment, DNA from the bacteriophage Lambda (48,502 base pairs in length) is cut with a variety of restriction enzymes and the resulting fragments are separated using gel electrophoresis. Three samples of Lambda (phage) DNA are incubated at 37º C, each with one of the 3 restriction endonuclease enzymes: Pst1, EcoRI, and HindIII.
What is the sequence of bacteriophage lambda?
The sequence of bacteriophage Lambda DNA. Lambda is a medium size E.coli bacteriophage. The DNA molecule of 48502 basepairs is linear and except for the extreme ends double-stranded. At each end the 5′ strand overhangs the 3′ strand by 12 bases.
What is lambda DNA used for?
Lambda DNA (48,502 bp) may be used as a molecular weight size marker during nucleic acid gel analysis following digestion with a restriction enzyme (such as HindIII). Lambda DNA can also be used as a substrate in restriction enzyme activity assays. Cat. #
What causes the overhang at the COS-ends of lambda DNA?
The 12 nucleotide 5’ overhangs at the cos-ends of the linear Lambda DNA are the result of a cut by the enzyme terminase. This enzyme is encoded by Lambda itself and acts like a restriction enzyme during the replication of the phage DNA. It is an endonuclease specific for the cos-site in multimeric phage DNA.