DNA, Genetics, Evolution Flashcards
Differences between DNA and RNA
DNA structure, Crick and Watson, Rosalind Franklin
- DNA molecular structure correctly proposed by James Watson & Francis Crick.
- They constructed models to quickly visualise & assess viability of potential structures
- 1st model was triple helix with bases on outside & sugar-phosphate residues in centre, with Mg cross-links between strands.
- Guided by:
- Molecular distances & bond angles
– Linus Pauling - DNA composed of nucleotides made up of sugar + phosphate + base.
– Phoebus Levene -
DNA X-ray crystallography data showed it’s organised into helical structure – RF
(without permission):- DNA purified, then fibres were stretched in thin glass tube (to make most strands parallel)
- DNA targeted by X-ray beam, which diffracts when it contacted an atom
- Scattering pattern of X-ray distinctive
so recorded on film & used to find
DNA molecular structure: -
Structure:
- X in centre of diffraction pattern indicated DNA = double helix.
- DNA molecule shown to twist at regular intervals to form helix.
- X-shape angle showed pitch of helix.
- Orientation: N-bases closely packed together on inside & phosphates form an outer backbone
- Molecular distances & bond angles
- Rosalind Franklin rejected model as not enough Mg & didn’t support Erwin Chargaff’s findings (# of A + G = # of T + C).
- Using trial & error, Watson & Crick assembled DNA model that showed:
- DNA strands antiparallel & form double helix.
- DNA strands pair via comp. base pairing (A = T ; C Ξ G)
- Outer edges of bases remain exposed (allows access to proteins involved in transcription & replication)
- Potential DNA replication mechansims:
- Replication occurs via comp. base pairing (A pairs with T, G pairs with C)
- Replication is bi-directional due to antiparallel nature of strands
DNA Replication
Context:
- Occurs during S phase of Interphase.
- Replication is a semi-conservative process as new DNA contains 1 old & 1 new strand.
- Parent DNA strands act as templates for new
strands. - Comp. bases from opp. strands form H-bonds (A = T & C ≡ G)
- Occurs in a 5’ to 3’ direction.
- Consequently, when DNA replicated:
- Each new strand formed identical to original strand separated from template
- Both DNA molecules formed have equal
base seq. to original molecule.
DNA replication Process
- Helicase: Unwinds double helix by breaking H-bonds between 2 strands. → forms replic. fork with strands running in antiparallel directions.
- DNA Gyrase: ↓ strain created by helicase by relaxing + supercoils (via (–) supercoiling).
-
Single Stranded Binding (SSB) Proteins:
- Split DNA strands & prevent strand from re-annealing
- Prevent nucleases from digesting SS-DNA
- Dislodged from strand when new comp. strand synth. by DNA pol III.
- RNA Primase: Synthesises RNA primers on each template strand, RNA primers are binding spots for DNA-PIII.
-
DNA-PIII: Once bound to primer:
- Cleaves PO4’s from nearby DNsTP, to form nucleotides which are added to 3’ end of primer; according to comp. base pairing
(A = T + C ≡ G) - Cleaving PO4’s releases nrg used to form covalent bond between nucleotides.
- Thus, DNA-PIII synthesises new DNA between RNA primers (Okazaki frags).
- Cleaves PO4’s from nearby DNsTP, to form nucleotides which are added to 3’ end of primer; according to comp. base pairing
- Leading Strand: Con. synth. where DNA-PIII moves towards replic. fork.
- Lagging Strand: Discon. synth. where DNA-PIII moves away from fork → create Okazaki frags as fork exposes more temp. strand.
- DNA-PI: Excises RNA primers & replaces them with DNA;
- DNA ligase: Joins gaps between Okazaki frags by making cov. bonds between nucleotides to form a continuous strand.
Transcription
Sections of Gene:
- Promoter: Non-coding seq. that acts as binding site for RNA pol, thus starting transcription.
- Sense Strand: DNA strand with same base sequence as RNA. Not transcribed.
- Antisense Strand: Temp. strand with comp. base seq. to RNA & sense strand. Transcribed.
- Terminator: Non-coding seq. that signals RNA pol to detach from DNA, thus ends transcript.
Process:
- Transcription: RNA seq. synthesis using DNA template, by RNA polymerase. Occurs in nuc.
-
Initiation:
- RNA pol binds to promoter & unwinds + separates DNA strands by breaking H-bonds between comp. base pairs (using NRG).
- Nucleoside triphosphates (NsTP) line up opp. their comp. base partner.
-
Elongation:
- RNA pol excises 2 phosphates → NRG
- NRG used to bind (now) free nucleotides together (thus, synth. RNA) in 5’ → 3’ dir.
- (Forms coding seq.)
-
Termination:
-
RNA pol reaches terminator → both RNA pol & synth. RNA strand detach &
DNA double helix re-forms.
-
RNA pol reaches terminator → both RNA pol & synth. RNA strand detach &
Nucleosomes
- In eukaryotic organisms, DNA packaged with histones to form (nucleosome).
- Histones: Proteins used by cell to package DNA into nucleosomes.
-
Nucleosomes: Molecules consisting of 8 histones (octamer) with DNA coiled around.
- Help to supercoil DNA, resulting in greatly compacted structure that allows for more efficient storage.
- Histone tails are + charged, so associate to DNA & determine tightness of packing
-
Acetylation: Adding acetyl group to histone tail, dec. + charge → DNA less tightly coiled & inc. transcript.
(Euchromatin) -
Methylation: Adding methyl group to histone tail, inc. + charge → DNA more tightly coiled & dec. transcript.
(Heterochromatin)
-
Acetylation: Adding acetyl group to histone tail, dec. + charge → DNA less tightly coiled & inc. transcript.
- Supercoiling helps:
- Protect DNA from damage
- Mobilise chrom’s during mitosis & meiosis.
Organisation of Eukaryotic DNA
- DNA packaged with 8 histone proteins (an octamer) → complex (nucleosome)
- H1 histone binds to linker DNA, which binds nucleosomes together, to form
* *chromatosomes** - These coil to form more condensed solenoid fibre structure, which then form loops.
- Loops compressed & folded around protein scaffold to form chromatin (eu- or hetero-).
- Chromatin then supercoils during cell division to form chromosomes visible (when stained) under microscope
Epigenetics
Epigenetics: Study of phenotypic changes caused by variations in gene expression levels.
- DNA Methylation prevents TF binding, so ↓ gene expression/transcription.
- Thus, transcription of gene ind. prop. to DNA methyl.
- Epigenetic analysis shows that DNA methylation patterns/gene expression may change over course of a lifetime
- Diff cell types in same organism may have markedly diff DNA methylation patterns.
- Env. factors (e.g. diet, pathogen exposure, etc.) also affect DNA methyl. within cells.
- Also influenced by heritability but not genetically pre-determined, so identical twins may have diff DNA methyl. patterns.
Transcription Regulation
- Transcription regulated by 2 groups of proteins that mediate binding of RNA pol to promoter:
- Transcription factors (TF) form complex with RNA pol at promoter & don’t allow initiation without factors, hence their levels regulate gene expression.
-
Regulatory proteins bind to non-coding DNA seq. outside of promoter & interact with transcription factors:
- Activator proteins bind to enhancer
seq. & ↑ transcript. rate
(by mediating complex formation) - Repressor proteins bind to silencer
seq. & ↓ transcription rate
(by preventing complex formation)
- Activator proteins bind to enhancer
-
Control Elements: Exist in large amounts to further tighten control & coordination.
- Distal control elements bind to regulatory proteins.
- Proximal control elements bind to transcription factors.
- Presence of certain transcription factors or regulatory proteins may be tissue-specific
- Intracellular chem signals may also trigger change in [reg. prots] or [TF] in response to stimuli → gene expression changes in response to changes in conditions in/out cell:
- Humans produce different amounts of melanin depending on light exposure
- Morphogens: Uneven distr. in embryo & contribute to diff. gene express. patterns depending on their conc.
- Mutant allele “cs” in “C” gene in Siamese cats only produces tyosinase (pigment production) at < body temp.
- In eukaryotes, post-transcriptional mod. of transcript mRNA needed to form mature mRNA
- Ribosomes also separated from genetic material (DNA & RNA) by nucleus, so gen. needs to be moved.
3 post-transcriptional events:
-
Capping: Involves addition of methyl group to 5’-end of transcribed RNA
- Methylated cap provides protection against degradation by exonucleases
- Allows transcript to be recognised by ribosome.
-
Polyadenylation: Addition of poly-A tail to 3’-end of mRNA. (NOTE: A stands for adenine)
- Poly-A tail improves RNA transcript stability
- Facilitates its export from nucleus.
- Thus, mRNA mods allow ribosome to it out of nuc. (via nuclear pores) before transl.
-
Splicing: Removing introns from mRNA transcript.
- Within eukaryotic genes exist:
- Introns: Non-coding seq. which must be removed prior to forming mature mRNA.
-
Exons: Coding regions, which fuse
together when introns removed to form continuous seq.
-
Alternative splicing: Removing specific
exons → Gene seq. make diff polypep’s.
- Within eukaryotic genes exist:
-
Polysome: Group of ribosomes translating an mRNA seq. simul.
- In prok. they couple transl. + transc. due to no comp. & both occuring in 5’ → 3’.
Translation
Context:
- Translation: Polypeptide synthesis using base sequence of mRNA molecules (in ribosomes).
- mRNA seq. read by ribosome in base triplets (codons). Each codon codes for 1 AA.
- Gen code degenerate as >1 codon can code for same AA. Also allows silent mutations to occur, whereby a change in DNA seq. doesn’t alter polypeptide seq.
- Thus, order of codons in mRNA seq. determine AA order in polypeptide chain.
- 64 codon possibilities (4 bases 3 bases/codon)
- 3 components work together to synthesise polypeptides by translation:
- mRNA has sequence of codons that determines AA sequence of polypeptide.
- tRNAs have anticodons that bind to comp. codon on mRNA; they carry AA corresponding to that codon.
- Ribosomes are mRNA & tRNA binding sites; also catalyse polypeptide assembly.
Process:
- Initiation: Assembly of components that carry out translation (mRNA, tRNA, ribosome).
- Next, appropriate tRNA molecule binds to
codon via its anticodon (according to comp. base pairing) - Finally, large ribosomal subunit aligns itself to the tRNA molecule at the P site and forms a complex with the small subunit
- Ribosome composed of 2 sub-units:
- Small subunit binds to mRNA & moves along it until reaching start codon (AUG).
-
Large Subunit containing 3 tRNA binding sites: A, P, and E binds to small subunit.
* Elongation:
- Initiator tRNA (with methionine) binds to start codon “AUG” in P site of large sub.
- 2nd tRNA (with anticodon comp. to 2nd codon) binds to 2nd codon in A site of large subunit.
(max 2 tRNAs bound at once). - Ribosome catalyses pep bond between AAs in A & P site via condensation reactions → dipep.
- tRNA in P site now deacylated (no AA), whilst
tRNA in A site carries dipeptide. - Ribosome translocates 3 bases along mRNA in 5’ to 3’ dir, so 1st tRNA moves from P to E site, releasing it.
- 2nd tRNA takes place of 1st. so moves from A to P site, freeing A site.
- 3rd tRNA binds with anticodon comp. to 3rd
codon on mRNA in vacant A site. - Process repeated until stop codon is reached.
- Termination: Disassembly of components & release of polypep chain.
- Non-coding stop codon reached, which release factor signalling transl. end; polypep released.
- Ribosome disassembles back into its 2 indep. subunits.
Activating tRNA
Context:
- tRNA activation occurs in cytoplasm via tRNA-activating enzyme, tRNA, AA and ATP.
- Each AA recognised by specific enzyme
- But multiple tRNA molecules recognised by enzyme due to degeneracy.
Process:
- tAE binds to specific AA & ATP.
- Enzyme catalyses ATP hydrolysis → AMP + 2P
- AA binds to AMP → AA-AMP complex, linked by high energy bond, 2P is released.
- Bond act as energy store to provide most of
nrg used to make pep. bond during transl. - tRNA binds to tAE.
- AA cov. bonded to 3’ terminal of tRNA, releasing AMP attached to enzyme.
- tRNA molecule now “activated” & released.
Insulin production in bacteria
Context:
- Diabetes type II due to destruction of ß-cells that secrete insulin (hormone).
- Used to be treated with insulin produced from other animals (e.g. pigs) as they bind to human insulin receptor, but found to cause allergies.
- Genetic code = universal as same codons code for same AAs in all living things, gen. info
transferrable between species - Ability to transfer genes between species used to produce human insulin in bacteria (for mass production), with exactly same AA seq. as gene
transcribed & translated in human cells.
Process
- Desired gene seq. obtained by either:
- DNA isolated from cells & nuclei by centrifugation (heavy cell organelles sink).
- Using rev. transcriptase to convert mRNA → dDNA.
- Interest gene specifically amplified via PCR
- Plasmids used as vectors as they can auton. self-replicate & express genes
- Gene + plasmid cut with same restriction endo.
at specific recognition sites by cleaving sugar-phosphate backbone to create “sticky ends” - Gene now binds to plasmid as sticky ends of gene & vector overlap via comp. base pairing.
- Gene & vector spliced together by DNA ligase (which fuses their backbones together with phosphodiester bond) to form recomb DNA.
- Recomb. DNA introduced into host cell/org.
(transfection if prok or transformation if euk). - Antibiotic selection commonly to ID which cells have successfully incorporated recomb. DNA.
- Transgenic cells, once isolated & purified, express desired trait encoded by int. gene, so placed in fermenter to reproduce lots.
- (e.g. Insulin) produced, purified and sold for use (e.g. in diabetics).
Hershey-Chase Experiment
- Proteins & nucleic acids believed to be involved in composition of genetic material.
- Alfred Hershey & Martha Chase conducted a series of experiments to prove DNA was gen material (not protein).
- Known that viruses (E.g. T2 Bacteriophage)
consisted solely of DNA & protein coat and could transfer their genetic material into hosts. - T2’s were grown in 1 of 2 isotopic mediums in order to radioactively label specific viral part.
- Viruses grown in radioactive S (35S) had radiolabelled proteins (S present in proteins but not DNA)
- Viruses grown in radioactive P (32P) had radiolabelled DNA (P present in DNA but not proteins)
- Viruses allowed to infect bacterium (E. coli).
- Virus & bacteria separated via centrifugation
- Larger bacteria forms solid pellet whilst smaller viruses remains in supernatant
- So when pellet found to be radioactive when infected by 32P–viruses (DNA) but not 35S–viruses (protein), showed DNA passed on.
- Showed DNA, not protein, was gen material as DNA was transferred to bacteria.
Meselson and Stahl
- Prior to experiment, 3 hypotheses had been proposed for the method of replication of DNA:
- Conservative Model: Entirely new molecule is synthesised from a DNA template (which remains unaltered)
- Semi-Conservative Model: Each new molecule consists of 1 newly synthesised strand & 1 template strand
- Dispersive Model: New molecules made of segments of new & old DNA
- Meselson & Stahl experimentally tested validity of these models using N15 (heavier radioactive isotope of N14), an element present in bases).
- DNA cultured in N15 for many gens to ensure N15 was only N source in DNA, then transferred to, & induced to replicate in N14-only medium.
- DNA samples separated via centrifugation to find DNA composition in replica. molecules.
- DNA detected as it absorbs UV, hence creating dark band when tubes illuminated with UV:
- Single band in 1st gen falsifies cons. replica. (shows mix of old & new DNA/
N15 & N14). - 2 bands in 2nd gen falsifies dispersive replication (New-only & mixed DNA /
Only-N14 and N15 & N14).
- Single band in 1st gen falsifies cons. replica. (shows mix of old & new DNA/
- Hence, showing DNA Replication is semi-conservative.
DNA Sequencing
- DNA sequencing: Process by which base order of a nucleotide sequence is elucidated
-
Dideoxynucleotides (ddNT): Lack 3’-OH
group needed for making PPD bond.- Thus, ddNTs stop further elongation of nucleotide chain & effectively end replica.
- Resulting DNA seq. reflects specific nucleotide pos. ddNT was added.
-
Sanger Method:
- PCR mixes set up, each containing stocks of deoxyribonucleotides + ddNT + fluoresc. primers + enzymes for replica.
- PCR makes lots of DNA molecules quickly, so PCR mixes should have all possible terminating frags for that spec. base.
- Frags separated using gel electroph & base seq. determined by ordering frags according to length.
- Fuorescently labelled primer included in each mix allow frags to be detected by automated seq. machines.
- If Sanger method conducted on coding strand, resulting seq. elucidated will be identical to template strand.
Non-coding DNA
- Vast majority of human genome is comprised of non-coding DNA, which serve other functions (table).
Context:
- DNA profiling: Technique by which individuals identified & compared via resp. DNA profiles
- Within non-coding regions of individual’s genome exists satellite DNA: DNA seq. made up of repeating elements (STRs).
Process:
- DNA sample collected (e.g. from blood, semen, saliva, etc.), then amplified using PCR
- Sat. DNA (with STR seq.) cut with specific restriction endo. to create frags, which differ between indivs due to # of STRs in frags.
3. Frags separated using gel electrophoresis & resulting profiles (composed of bands) are compared to see if bands match.
-
For family tests:
- Paternal lineage determined by analysing VNTR from Y-chromosome.
- Maternal lineage deduced by analysing mitochondrial DNA variations in single nucleotides at hyper-variable regions.
Genes, Locii, Alleles, Chromosomes, Homologous Chromosomes, SNPs, Mutations,
- Polygenic traits: Traits influenced by multiple genes.
- Gene: Heritable factor consisting of a length of DNA and influences a specific characteristic.
- Locus: Specific position on chrom occupied by gene.
-
Allele: Alt. forms of same gene, they have same locus, but only one can occupy it.
- Code for diff. variations of specific trait
- Alleles have very similar gene seq. & only diff. by 1 or few bases (SNP’s).
- Single Nucleotide Polymorphisms (SNP): Positions in gene where >1 base may be present.
-
Chromosomes: Groups of linked genes.
23 types of chromosomes in humans.- Homologous: Chromosomes that carry same sequence of genes but not necessarily same alleles of those genes, allowing species members to interbreed.
-
Mutations: Random changes in base seq. of gene. May lead to new alleles forming from other alleles.
- Base Substitution: 1 base in seq. of gene replaced by different base.
- Almost all mutations either neutral or harmful as random change to allele selected for by NS over time unlikely to be beneficial.
- Mutations in body cells eliminated when individual dies, but mutations in sex cells can be passed on to offspring & cause genetic disease.
Mutations
- Gene mutation: Change in nucleotide/base seq. of gene coding for a specific trait.
- New alleles are formed by mutation
(alleles only differ by few bases (SNP’s)). - Either spontaneous or induced.
Gene mutations can be:
- Beneficial: Change gene seq. (missense mutations) to create new variations of a trait
-
Detrimental: Truncate gene seq. (nonsense
mutations) to STOP normal function of trait. - Neutral: Have no effect on functioning of specific trait (silent mutations).
Induced by:
- Ionising Radiation ↑ mutation rate if enough nrg to cause chemical changes in DNA.
-
Chemical Mutagens ↑ mutation rate by causing chemical changes in DNA.
(e. g. mustard gas, benzene, ROS, tar) - Biological: Viruses (HPV) and Bacteria
Hiroshima + Nagasaki and Chernobyl
Sickle Cell Anaemia
- Disorder caused by single base sub. mutation to a gene (Hb), which codes for haemog. prod.
- Most humans have co-dom. allele Hb4.
-
DNA: CTC → CAC on 6th codon of gene
→ new co-dom. allele formed: Hb5. - mRNA: GAG → GUG.
- Polypeptide: 6th AA in haemog: Glu → Val when GUG transcribed.
- Mutation → changes haemog. structure →
stick to form insoluble fibrous strands → rigid enough to distort RBCs into sickle shape. -
Consequences:
- Damage to tissues by becoming trapped in & blocking blood capills → ↓ bloodflow.
- Both haemog & plasma memb. dmg
- RBC Life ↓ → ↓ RBC count → Body can’t replace RBC at same rate → anaemia.
Genome + HGP (PM MS MEGI)
Genome: Whole of gen. info of organelle, cell, or organism. Includes genes + non-coding DNA seq.
- In animals, genome = DNA molecules that form chromosomes in nucleus + DNA molecule in mitochondrion.
- 46 chromosomes in humans.
- In plants, genome = DNA molecules that form chromosomes in nucleus + DNA molecules in mitochondrion + chloroplast.
- In prokaryotes, genome = DNA in circular chromosome + any plasmids that are present. Much smaller as a result.
- Genome size generally ∝ size:
- Viruses & bacteria tend to have smallest genomes
- Plant genome size varies dramatically due to capacity for plant species to self-fertilise & become polyploid.
- Size may also change due to chromosomes fusing or splitting, but rare.
HGP: International cooperative venture established to sequence human genome
- HGP showed that humans share majority of their seq, with SNP’s contributing diversity.
- HGP aided by improvements in tech that rapidly inc. speed of gene sequencing.
- Completion of HGP led to many outcomes:
- Mapping: #, location, size & seq. of human genes now established
- Screening: Allowed for prod. of specific gene probes to detect sufferers & carriers of genetic diseases.
- Medicine: Discovery of new proteins & causes of gen. diseases → ↑ treatments.
- Evolution: Comparing with other genomes → ↑ knowledge of origins, evolution & migratory patterns of man
- Gene transfer (genetic engineering)
- Promote International co-operation.
- Understanding that genome > proteome & that most genome not transcribed.
- Mutations discovered.
(e.g. introns, promoters, STR’s, etc.)
Euk + Prok DNA + Plasmids
-
Plasmids:
- Contain few genes
- Capable of self-replication
- Exchanged between bacteria via pili (conjugation) → bacteria evolves new features within gen (horiz. gene transfer).
- Plasmids may also cross species barriers if plasmid released when prok. absorbed by cell of different species.
- Plasmid’s ability to self-replicate & synth. proteins → vectors for genetic engineering.
Homologous Pairs
- Sexually reproducing organisms inherit their genetic seq. from both parents
- So organisms possess 2 copies of each chrom (homologous chrom), which share:
- Same structural features (e.g. same size, same banding patterns, same centromere positions)
- Same genes at same loci positions (whilst
genes are same, alleles may be different) - Homo. chrom. separated in gametes (via meiosis) prior to reproduction, in order to prevent chrom. numbers continually doubling with each generation.
- Organisms with diff. diploid #’s unlikely to be able to interbreed (can’t form homo. pairs in zygotes).
- In cases where diff. species do interbreed, offspring usually infertile (can’t form functional gametes). (e.g. horse + donkey).
Karyograms
-
Karyotypes: # & types of chrom. in euk:
- Harvest cells (usually from foetus or adult WBC’s).
- Cell div chemically induced, then mitosis arrested whilst chrom. are condensed
(so they’re visible). - Stage during which mitosis halted determines whether chrom. appear with sister chromatids or not.
- Chroms stained & photo taken, then
arranged into homo. pairs by size
(sex chrom. shown last) → karyogram.
-
Karyotyping usually done prenatally to:
- Determine gender of unborn child (via ID of sex chrom.)
- Test for chromosomal abnormalities (e.g. Down)
-
Down syndrome:
- Due to non-disjunction in 1 of parental gametes: Failure of chrom separation resulting in 1 xtra/1 less chrom.
- Non-disjunction may occur via:
- Sis chromatids in Anaphase II
→ 2 affected daughter cells - Bivs failing to separate in Anaphase I → 4 affected daughter cells.
- Sis chromatids in Anaphase II
- Down = Trisomy due to: 1 parental gamete having 2 chrom 21 copies (due to non-disj) + 1 normal parental gamete with 1 copy fusing → 3-copy zygote.
- Studies show that:
- Non-disj ∝ parental (esp. mom) age ↑
- May be due to developing oocytes being arrested in prophase I until ovulation as part of oogenesis.
- Higher incidence of chrom. errors in offspring due to anaphase I non-disjunction.
- Mean maternal age ↑, → ↑
# of Down syndrome offspring.
- Non-disj ∝ parental (esp. mom) age ↑
-
Karyotyping sources:
- Amniocentesis: Extraction of amniotic fluid (has fetal cells) with needle inserted through abdomen.
-
Chorionic Villus Sampling (CVS):
Extraction of CV (placental tissue) with suction tube inserted through cervix. - CVS can be done earlier in preg. than
Amniocentesis, but risk = 2% (opp. to 1%).
Autoradiography
John Cairn 1st to create image of chroms before condensation, which made measuring inaccurate.
- Incubate cells in radioactive 3H-T solution.
- 3H-T incorporated into chromosomal DNA of cell as T not present in RNA.
- Chromosomes isolated by gently lysing cells & fixing chrom. to photographic surface
- Surface then immersed in radioactively-sensitive emulsion containing AgBr.
- Radiation released from 3H-T converts Ag+ ions in AgBr into insoluble Ag grains.
- Following period of exposure, excess AgBr
washed away, leaving Ag grains = small black • - When photographic film developed, chrom. DNA visualised with an electron microscope.